Browsing by Subject "eläinlääketiede"

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  • Orro, Toomas (Helsingin yliopisto, 2008)
    The early protection mechanism of the host against infection, trauma or other tissue damage comprises a set of reactions known as the acute phase response (APR). During APR, circulating concentrations of acute phase proteins (APPs) change. These proteins can serve as indicators of host response during various inflammatory conditions. In this thesis, APR in reindeer was investigated for the first time. Systemic concentrations of APPs during the neonatal period were studied in reindeer and cattle. APPs were also investigated during spontaneous bovine respiratory disease (BRD) in dairy calves. Escherichia coli endotoxin challenge in adult reindeer increased concentrations of serum amyloid A (SAA) in all animals. Haptoglobin (Hp) showed a less pronounced increase. SAA and Hp were concluded to be acute phase reactants in reindeer. In reindeer calves SAA concentrations increased during the first 2 weeks of life and decreased afterwards. Serum Hp concentrations increased throughout the first month after birth. In dairy calves SAA and lipopolysaccharide binding protein (LBP) concentrations changed similarly during first month of age as in reindeer calves. However, Hp concentrations generally remained low after birth. SAA rise in calves were not derived from colostrum as mammary specific SAA isoforms were not found from calves serum samples. Results of these two studies indicated that newborn reindeer and dairy calves have an inflammatory response during the first weeks of life and the age of young animals should by considered when interpreting APP concentrations. Very similar SAA changes in the two ruminant species also suggest that this inflammatory response may have role in the adaptation process of newborns to extrauterine life. The effect of different bovine respiratory pathogens on concentrations of APPs (SAA, LBP, Hp, alpha1-acid glycoprotein and fibrinogen) was studied in calves. Isolation of Pasteurella multocida was associated with increased concentrations of all APPs tested. In another study, concentrations of APPs were investigated in dairy calves during an outbreak of BRD. Initial cause for BRD outbreak was bovine respiratory syncytial virus (BRSV) infection. Concentrations of SAA and LBP increased in parallel with clinical symptoms at week 1 and peaked at week 3 of outbreak. Some calves had high Hp concentrations at week 3. Higher SAA, LBP and Hp concentrations at a later stage of BRD (week 3) were associated with lower BRSV-specific IgG1 production, suggesting that these calves had enhanced inflammatory response to secondary bacterial infection. In conclusion, APPs proved to be useful in exploring host response in bovine respiratory infections.
  • Herva, Tuomas (Helsingin yliopisto, 2015)
    Animal welfare (AW) is an issue of growing concern in Finland as well as in other developed countries. A public debate has focused on the potential AW problems resulting from current production systems. Possibilities to find mutual benefit for animals, farmers, industry and society have received less attention. According to the reviewed literature the inconsistency of determination and perception of AW appeared to be a major barrier to enhance AW. Farmers should be confident that their measures to promote AW satisfy public opinion and are ecomically sustainable. The main objective of the study was a thorough understanding of relationships between AW and beef production economics to find barriers and opportunities for enhanced AW. A version of the Animal Needs Index (ANI/TGI 35L), modified for Finnish beef production, called A-index was used for AW assessments. The A-Index was modified and evaluated based on Test Theory. On-field associations between A-index and production parameters were determined on 180 farms and over 12 000 bulls using statistical multilevel models. Economic evaluation of AW was based on comparison between cold and warm housing using the confirmed association between AW and production results. AW was associated with good production results. A-Index and the best subset of items used as welfare score (WFS) were covering different aspects of AW. The association between the used measures and production results, reflecting AW in certain degree, can be considered as a proof of the criterion validity of A-Index and WFS. Cold housing with enhanced welfare and bedding based on own straw at a reasonable price was economically favourable. Profitability of cold housing was sensitive to fluctuation in bedding price. Developing a reasonably priced market for bedding material would be a major way to enhance AW. Rubber covered slats were found to be a profitable way to enhance AW in warm housing. A reform of the subsidy system was suggested to be needed to fulfil the aims of the subsidy regime to support AW.
  • Hielm-Björkman, Anna (Helsingin yliopisto, 2007)
    The series of investigations presented in this thesis examined different methods of assessing chronic pain in dogs suffering from osteoarthritis (OA) and compared the effects of three different treatments. Data were obtained from two cohorts; 41 dogs with OA due to canine hip dysplasia (CHD) (I,III) and 61 dogs with OA due to CHD or elbow dysplasia (II,IV,V). Questionnaires, veterinary evaluations, visual analog scales (VAS), plasma hormones, radiographs, and force plate evaluations were assessed as OA treatment outcome measures and/or measurements of chronic pain. The results indicated that the multidimensional pain scale including 11 questions with 5-point scale responses was a valid and reliable tool for evaluating chronic pain. This Helsinki chronic pain index (HCPI) can be applied as an outcome measure in clinical trials where chronic pain is evaluated by owners. Of the evaluated complementary therapies for chronic pain due to OA, all three indicated a positive treatment outcome. In the first trial, gold bead implants resulted in a significant positive treatment outcome for the treatment group. However, the placebo group in this study also improved significantly. A positive effect was seen in 65% of the placebo dogs and this exceptionally high incidence of amelioration suggests that the placebo group may have got an effect of unintentional needle acupuncture. The results of this study are therefore controversial and treatment guidelines based on these findings cannot be given. The second trial tested two ingestible OA remedies, green lipped mussel and a homeopathic low-dose combination preparation. Both treatments resulted in statistically significant positive treatment outcomes compared with placebo, but with the positive control (carprofen) being more effective than either of them. The results suggest that both tested treatments may be beneficial for chronic OA. To establish the true role of all these three treatments in outcome-based animal analgesia, more clinical trials, using larger cohorts, should be conducted. Possible mechanisms of action should also be studied.
  • Taponen, Suvi (Helsingin yliopisto, 2008)
    Bovine mastitis caused by coagulase-negative staphylococci Mastitis is the most common disease affecting dairy cows, and staphylococci are the most common cause of mastitis. According to Finnish studies, staphylococci are isolated from 36% of clinical and 60% of subclinical mastitis cases. In mastitis diagnostics, staphylococci are divided into coagulase-positive and coagulase-negative based on their ability to coagulate plasma. Majority of staphylococcal mastitis is caused by coagulase-negative staphylococci (CNS). In mastitis diagnostics they are not identified at species level but are treated as a uniform group. Usually they cause subclinical or mildly clinical mastitis, and are considered as harmless bacteria of the normal skin flora. CNS mastitis is most common on the first lactation. It is assumed to cure spontaneously and antimicrobial treatment is not recommended. Although usually subclinical or mildly clinical, CNS mastitis causes increase of milk leucocytes and may decrease the milk quality. CNS mastitis seems to be especially a problem in well managed dairy herds with good udder health. In contrast to the usual assumption that CNS mastitis cures spontaneously, it may persist in the udder throughout the entire lactation. In addition, the CNS species are not necessarily a uniform group but differences between the species in virulence and clinical characteristics may exist. In this dissertation, different aspects of bovine mastitis caused by CNS were studied. The response of CNS mastitis to antimicrobial treatment was investigated, as well as the persistence of CNS intramammary infection when left without antimicrobial treatment. The prevalence of different CNS species in mastitis and on cows skin, and possible differences in clinical characteristics and persistence between CNS species were studied. Species identification was performed based on bacterial phenotype and genotype. Identification using different methods was compared. Majority of CNS mastitis was caused by S. chromogenes or S. simulans. Many other CNS species were isolated from mastitis sporadically. Molecular genetic identification methods based on bacterial genotype proved more reliable than biochemical methods based on bacterial phenotype. In some cases the identification results of genotypic and phenotypic methods disagreed. About half of CNS mastitis detected during lactation persisted without antimicrobial treatment, commonly until the end of lactation. Mastitis caused by penicillin-susceptible CNS cured well, about 90% cured. Of mastitis caused by penicillin-resistant CNS, 60 to 70% cured, although the causing bacteria were susceptible to the antimicrobial used. Most often CNS mastitis was subclinical or mildly clinical. Statistically significant differences in clinical characteristics or persistence between the two most common CNS species were not detected.
  • Simojoki, Heli (Helsingin yliopisto, 2011)
    Coagulase-negative staphylococci (CNS) are the most common bacteria isolated in bovine subclinical mastitis in many countries, and also a frequent cause of clinical mastitis. The most common species isolated are Staphylococcus (S) chromogenes, S. simulans, S. epidermidis, and S. xylosus. One half of the intramammary infections (IMI) caused by CNS persist in the udder. The pathogenesis of IMI caused by CNS is poorly understood. This dissertation focuses on host response in experimental intramammary infection induced by S. chromogenes, S. epidermidis and S. simulans. Model for a mild experimental CNS infection was developed with S. chromogenes (study I). All cows were infected and most developed subclinical mastitis. In study II the innate immune response to S. epidermidis and S. simulans IMI was compared in eight cows using a crossover design. A larger dose of bacteria was used to induce clinical mastitis. All cows became infected and showed mild to moderate clinical signs of mastitis. S. simulans caused a slightly stronger innate immune response than S. epidermidis, with significantly higher concentrations of the interleukins IL-1beta and IL-8 in the milk. The spontaneous elimination rate of the 16 IMIs was 31%, with no difference between species. No significant differences were recorded between infections eliminated spontaneously or remaining persistent, although the response was stronger in IMIs eliminated spontaneously, except the concentration of TNF-α, which remained elevated in persistent infections. Lactoferrin (Lf) is a component of the humoral defence of the host and is present at low concentrations in the milk. The concentration of Lf in milk is high during the dry period, in colostrum, and in mastitic milk. The effect of an inherent, high concentration of Lf in the milk on experimental IMI induced with S. chromogenes was studied in transgenic cows that expressed recombinant human Lf in their milk. Human Lf did not prevent S. chromogenes IMI, but the host response was milder in transgenic cows than in normal cows, and the former eliminated infection faster. Biofilm production has been suggested to promote persistence of IMI. Phenotypic biofilm formation and slime producing ability of CNS isolates from bovine mastitis was investigated in vitro. One-third of mastitis isolates produced biofilm. Slime production was less frequent for isolates of the most common mastitis causing species S. chromogenes and S. simulans compared with S. epidermidis. No association was found between the phenotypic ability to form biofilm and the persistence of IMI or severity of mastitis. Slime production was associated with persistent infections, but only 8% of isolates produced slime.
  • Suojala, Leena (Helsingin yliopisto, 2010)
    Bovine mastitis caused by Escherichia coli remains a problem in many countries despite of improvements in managing and housing of dairy cattle. Cows with compromised immune systems, especially those in early lactation, are particularly susceptible to E. coli intramammary infection. Mastitis caused by E. coli varies from mild, with local signs only, to severe systemic. The severe form of E. coli mastitis is associated with loss of milk production and can result in death of the cow. Intramammary infection caused by E. coli is often eliminated spontaneously as the defense mechanisms of the cow are able to clear the bacteria from the udder. The aims of this thesis were to investigate host response to E. coli mastitis in terms of characteristics of bacteria, effect of antimicrobial and non-antimicrobial treatment, and prophylactic effect of recombinant human lactoferrin expressed in milk. Host response was studied using two consecutive intramammary challenges with E. coli in the same cows at a short interval. All cows became infected, but local signs were significantly milder and disappeared faster after the second challenge. The same pattern was recorded for the indicators of inflammation; the differences being statistically significant for serum and milk haptoglobin, milk serum amyloid A, and white blood cell count. Milk production returned to the pre-challenge level significantly faster after the second challenge. This study revealed a possible carry-over or immunizing effect of the previous intramammary infection by the same pathogen. Broad-spectrum antimicrobials have been widely used for treating E. coli mastitis, although the results of treatment studies have been controversial. In our field study, systemic enrofloxacin treatment did not result in better bacteriological or clinical cure of acute clinical mastitis caused by E. coli than supportive treatment alone. Enrofloxacin treatment did not affect survival of the cows, return of quarter milk production or tissue damage of the affected quarter within three weeks post-treatment; nor did it affect the length of time the cow remained in the herd during the six-month follow-up period. The only positive effect of the enrofloxacin treatment was a higher bacteriological cure 2 days post-treatment. On the contrary, clinical cure on day 2 was lower in the enrofloxacin treated cows. Clinical cure assessed 21 days post-treatment was relatively low in both groups, which probably reflects the severe nature of acute E. coli mastitis. Our study did not support the use of parenteral antimicrobial treatment of clinical E. coli mastitis. In severe cases of coliform mastitis, antimicrobial treatment could, however, still be recommended for safety reasons, as it can increase the elimination of bacteria and prevent possible bacteraemia. Our results indicated also that frequent milking at the acute stage of mastitis might improve the initial clinical cure of E. coli mastitis and decrease inflammation and tissue damage in the infected quarter. Most E. coli isolates belonged to the phylogenetic group A, indicating their commensal nature. Close to 40% of the isolates had at least one virulence gene, but combinations of virulence genes varied greatly, each combination being present mainly in a single isolate. Approximately 30% of the isolates showed resistance to one or more antimicrobials tested, most commonly against ampicillin, streptomycin, tetracycline and sulphonamides. No specific virulence factor, phylogenetic group or resistance to antimicrobials was associated with persistence or severity of disease among mastitis E. coli isolates in our field study. These results indicate that characteristics of bacteria are not likely to affect the clinical course and outcome of E. coli mastitis. In 11% of mastitis cases, the same genotype of E. coli was isolated from the affected quarter three weeks post-treatment as originally, indicating persistence of the same genotype of E. coli in the quarter. Non-antimicrobial treatments, such as lactoferrin, which has antibacterial and lipopolysaccharide neutralizing properties, could be beneficial in the treatment of E. coli mastitis. The efficacy of intramammary lactoferrin was compared with that of systemic enrofloxacin in an experimentally induced E. coli model. No significant differences were found in the clinical signs between cows treated with lactoferrin and those treated with enrofloxacin. The results from this study remained inconclusive. The prophylactic effect of lactoferrin against E. coli mastitis was studied in a new model of using hLf-transgenic cows, which expressed recombinant human lactoferrin in their milk. This was the first study to describe an experimentally induced E. coli mastitis model using transgenic cows. The high concentration of lactoferrin in the milk of the transgenic cows did not protect the cows from E. coli intramammary infection, and all of them became infected. No differences were noted in the bacterial growth, times to bacterial elimination, local signs or in any of the milk or blood inflammatory parameters, except in concentrations of haptoglobin and cortisol in the serum. However, systemic signs were significantly milder in transgenic cows. Prevention of E. coli mastitis relies on decreasing the infection pressure in the environment of the cows and improving the cow comfort and herd management. In E. coli mastitis, the treatment should be as efficient as possible, but novel therapeutic approaches are needed, as the efficacy of the current commonly used antimicrobial treatments is not satisfactory.
  • Perko-Mäkelä, Päivikki (Helsingin yliopisto, 2011)
    Campylobacter, mainly Campylobacter jejuni and C. coli, are worldwide recognized as a major cause of bacterial food-borne gastroenteritis. Epidemiological studies have shown handling or eating of poultry to be significant risk factors for human infections. Campylobacter contamination can occur at all stages of a poultry meat production cycle. The aim of this thesis was to study the occurrence and diversity of Campylobacter in broiler and turkey production in Finland. In summer 1999, 2.9 % of slaughtered broiler flocks were Campylobacter-positive. From the isolated strains 94 % were C. jejuni and 6% were C. coli. During years 2005-2006 one turkey parent flock, the hatchery, six different commercial turkey farms and different stages of the slaughterhouse were monitored during one and the half year. No Campylobacter were detected in either of the samples from the turkey parent flock or from the hatchery using the culture method. Instead PCR detected DNA of Campylobacter from the turkey parent flock and samples from the hatchery. Six out of 12 commercial turkey flocks were found negative at the farm level but only two of those were negative at slaughter. Campylobacter-positive samples within the flock at slaughter were detected between 0% and 94% with evisceration and chilling water being the most critical stages for contamination. All of Campylobacter isolates were shown to be C. jejuni. Campylobacter-positive turkey flocks were colonized by a limited number of Campylobacter genotypes both at the farm and slaughter level. In conclusion, in our first study in 1999 a low prevalence of Campylobacter in Finnish broiler flocks was detected and it has remained at a low level during the study period until the present. In the turkey meat production, we found that flocks which were negative at the farm became contaminated with Campylobacter at the slaughter process. These results suggest that proper and efficient cleaning and disinfection of slaughter and processing premises are needed to avoid cross-contamination. Prevention of colonization at the farm by a high level of biosecurity control and hygiene may be one of the most efficient ways to reduce the amount of Campylobacter-positive poultry meat in Finland. With a persistent low level of Campylobacter-positive flocks, it could be speculated that the use of logistic slaughtering, according to Campylobacter status at farm, might have be advantageous in reducing Campylobacter contamination of retail poultry products. However, the significance of the domestic poultry meat for human campylobacteriosis in Finland should be evaluated.
  • Rikula, Ulla Kaisa (Helsingin yliopisto, 2008)
    Canine distemper (CD) is one of the longest-known infectious diseases of dogs and many other carnivores. Control of CD among dogs relies heavily on vaccination, while in fur farms and zoos biosecurity measures in some cases offer the only available means for CD control. Modified live CD virus vaccines have been successfully used to control CD among farmed mink, and since no licensed vaccines for other species kept for fur exist, they have also been used for foxes and raccoon dogs in CD emergency situations. CD vaccines for dogs and mink were studied in experimental settings for their ability to induce virus-neutralising (VN) antibodies in target species. Mink vaccines were also assessed in silver foxes, blue foxes and raccoon dogs. Purpose-bred beagle dogs were vaccinated twice with one of three CD vaccines: Candur® SHP, Canlan®-3 or Dohyvac® DA2P, and the levels of VN antibodies were determined at the time of vaccination and one month after the second vaccination. Fur animals were vaccinated once with Distemink®, Distem®-R-TC or vaccine 3 and the levels of VN antibodies were determined at vaccination and 2-4 times 1-4 months afterwards. Significant differences among vaccine groups were found both in the proportion of animals with measurable levels of VN antibodies and in the mean titres of antibodies. The levels of VN antibodies were also determined from a large field sample (n = 4 627) of vaccinated dogs. In addition to the three CD vaccines in the seroconversion study above, additional two vaccines, Duramune®-4 and Nobivac® DHP, had been used in the field. Each dog with a known vaccination history, date of birth, sex and breed was sampled once. Based on the overall geometric mean titre of the dogs vaccinated with a single vaccine brand, vaccines were divided into high-take (Candur®, Nobivac® and Duramune®) and low-take (Dohyvac® and Canlan®) groups. The vaccine groups differed significantly among dogs less than two years of age both in the proportion of dogs with detectable VN antibodies and in the mean titres. Both the number of vaccinations and age were associated with the titre and vaccine usage. To control for possible confounding factors, the comparison of titres among vaccine usage groups was adjusted by classifying them according to the number of vaccinations (one to four) and the age group (less than one, one to two, or over two years old). The same division into low- and high-take vaccines was observed, irrespective of the number of vaccinations the dogs had received. The observations of this seroprevalence study regarding Candur®, Canlan® and Dohyvac® were consistent with the results of the seroconversion study. CD was reintroduced into Finland in 1990 after 16 years of absence. The disease remained at a low endemic level in 1990-1994, reached epidemic proportions in 1994-1995 and disappeared during 1995. The epidemic also involved vaccinated dogs. Among the virologically-confirmed cases the proportion of Dohyvac®-vaccinated dogs was higher than expected from the market shares on the assumption that all the vaccines had an equal take. As a result of this observation, Dohyvac® was withdrawn from and Nobivac® and Duramune® introduced to the market during 1995. A drastic redistribution of the market shares between the low-take and high-take vaccines took place, and this coincided with the decline and dying out of the outbreak. The observed occurrence pattern of CD from 1990-1996 was largely attributed to the changes in the level of HI, although the possible contribution of other factors, such as developments in the dog demographics, was also recognized. It was concluded that an HI above 75% is needed to keep CD in check, i.e., only sporadic cases of CD, at most, can occur. With the currently used vaccines an HI of 80% corresponds to a vaccine coverage of some 94%. It was concluded that the development of vaccine-induced immunity is a multifactorial process depending on the properties of the vaccine, on the individual variation, age, species and other factors influencing the immunocompetence of the host. On the individual level the prevention of clinical signs is sufficient, but on the population level, halting the circulation of the virus is crucial for the definitive control of CD. The ultimate test and criterion for a vaccine is its contribution to herd immunity.
  • Lilja-Maula, Liisa (Helsingin yliopisto, 2014)
    Canine idiopathic pulmonary fibrosis (CIPF) is an incurable fibrosing lung disease occurring mainly in West Highland White Terriers (WHWTs). The clinical picture of CIPF has many similarities with human idiopathic pulmonary fibrosis (IPF). Signs include dry cough, exercise intolerance, and respiratory difficulties. Prognosis for CIPF and human IPF is poor, and only limited treatment options are available. Histopathological CIPF shares features of both human usual interstitial pneumonia (UIP), the pathological counterpart of human IPF, and other human idiopathic interstitial pneumonia, the non-specific interstitial pneumonia (NSIP). Chronic bronchitis (CB) is the main differential diagnosis for CIPF, but antemortem differentiation can be challenging, as surgical lung biopsies are seldom taken and no clinically useful biomarkers are currently available. The natural history of CIPF has not been previously studied and little is known about the molecular pathophysiology of CIPF. This thesis describes the clinical course, survival, and evaluation of exercise intolerance using the 6-minute walk test in CIPF WHWTs. Bronchoalveolar lavage fluid (BALF) protein expression was studied to find potential biomarkers for CIPF, and aspects of transforming growth factor (TGF)-β related molecular pathways in pathogenesis of CIPF were investigated. In addition, results of TGF-β signaling activity and its known extracellular matrix (ECM) regulatory proteins, latent TGF-β binding protein (LTBP)-1 and fibrillin-2, in CIPF were compared with findings in human IPF/UIP and NSIP. The follow-up study showed that CIPF has a significant negative impact on life expectancy of diseased dogs. The median CIPF-specific survival after onset of clinical signs in WHWTs was 2.7 years, but individual survival varied considerably from only a few months to over 4 years. This variance indicates that CIPF may have either a slow or a rapid disease progression, as also seen in human IPF. In addition, during the disease course acute exacerbations (AEs) occurred. The 6-minute walk distance proved to be an easy and noninvasive parameter to evaluate lung function and level of exercise intolerance in CIPF WHWTs. No significant prognostic factors were identified. The quantitative comparison of BALF proteomes obtained from CIPF WHWTs, CB dogs, and healthy dogs revealed similar changes for CIPF and CB, which suggests a common response to disease processes in these otherwise different lung diseases. Specific biomarkers for CIPF were not identified. The immunohistochemical stainings suggested that increased TGF-β signaling and its ECM regulatory proteins LTBP-1 and fibrillin-2 are part of the molecular pathophysiology of CIPF, as also seen in human IPF/UIP and NSIP. We demonstrated strong expression of activin B, a member of the TGF-β superfamily, in the altered alveolar epithelium of WHWTs with CIPF. Furthermore, activin B was detected in BALF of CIPF WHWTs, most notably in samples from dogs with AE, but not in BALF of healthy WHWTs. This novel finding suggests that activin B is part of the CIPF pathophysiology and might act as a potential marker of alveolar epithelial damage. Our findings add important new knowledge about the natural history and molecular pathophysiology of CIPF in WHWTs and show similarities between CIPF and human IPF. Better understanding of the complex pathogenesis of CIPF and human IPF is crucial for developing novel diagnostic tools and treatment strategies for these yet incurable diseases.
  • Ahonen, Saija (Helsingin yliopisto, 2014)
    The availability of the genome sequence and genomic tools combined with the unique phenotypic diversity and breed structure has increased interest in dog as a large, physiologically relevant animal model for human genetic research. Dogs suffer from hundreds of hereditary conditions of which many resemble closely disorders in humans. Investigation of the genetic background of these conditions is important to understand the disease mechanisms and pathways for therapeutic options, to identify novel candidate genes for corresponding human conditions, and to inform the breeding programs. Remarkable resources have been established in the University of Helsinki for canine genetic research, including one of the largest canine DNA banks. This study has focused on degenerative eye diseases of unknown genetic cause. We combined efforts in basic and clinical veterinary research with a unique network of collaboration with dog breeders and owners to establish clinically relevant study cohorts for two types of retinal degeneration in Papillon and Phaléne, and in Swedish Vallhund, SV breeds and two different types of glaucoma in Dandie Dinmont Terriers, DDT and Norwegian Elkhounds, NE. Genome wide approaches with the latest genomic tools, including SNP arrays and exome sequencing were then utilized to identify causative loci and genes. We report breakthroughs in all projects and highlight shared molecular etiologies in canine and human vision disorders. The cause of the retinal degeneration in Papillons and Phalénes was found in the CNGB1 gene, which is important for the rod cell function and has been linked before to human retinal degenerations. A unique type of multifocal retinal degeneration was described in SV with association to the upregulation of the MERTK gene. MERTK mutations cause also retinal degeneration in humans. DDTs were found to present a primary closed angle glaucoma (PACG) with severe pectinate ligament dysplasia (PLD). The disease was mapped to a region on canine chromosome 8, which is syntenic to human chromosome 14 with multiple glaucoma loci. However, further studies are required to identify the causative mutation. Finally, we found that the primary glaucoma in NEs is caused by a missense mutation in the ADAMTS10 gene. This gene has been associated with a Weill-Marchesani syndrome (WMS) with ocular and non-ocular abnormalities in humans. This study provides new candidate genes, and mechanisms for human eye disorders, and has identified models for potential therapeutic trials. At the same time breeding programs benefit from novel gene tests to eradicate the blinding conditions from the studied breeds.
  • Gonzalez, Manuel (2012)
    Campylobacter Spp are recognized as a major cause of bacterial food-borne gastroenteritis worldwide, with Campylobacter jejuni and Campylobacter coli being the most common species isolated in human infections (WHO, 2011). The number of registered cases of human campylobacteriosis in Finland has ranged from 3,796 cases in 2001 to 4,231 cases in 2011. The reported incidence in Finland in the last 10 years is higher than the European Union average. In order to compare human, chicken and cattle C. jejuni isolates, the presence or absence of four nonubiquitous genes were determined so that they could be associated with the source of the isolate. First, we tested the presence of dmsA, which encodes a subunit of the putative tripartite anaerobic dimethyl sulfoxide oxidoreductase (DMSO/trimethylamine N-oxide reductase). Second, we detected cj1585c, which encodes another oxidoreductase. Third, the serine protease gene cjj81176-1367/1371 was isolated. Fourth, γ-glutamyl-transpeptidase gene ggt was detected. We ascertained that ggt and dmsA are present more frequently in isolates obtained from humans and chickens, whereas cjj81176-1367/1371 and cj1585c are the most common in bovine isolates. Campylobacter jejuni is able to survive in different environments and in a wide range of temperatures. The study of C. jejuni inactivation in minced chicken meat and dug well water ascertain that the Weibull model could be applied optimally to the data to build a reliable prediction model for the survival of this microorganism as a function of temperature. The longest survival time found for C. jejuni in minced meat chicken was at the storage temperature of -20°C, and that of dug well water was at 4°C. We analyzed the effect of different seasoning as dry marinade combinations on accelerating the reduction of C. jejuni counts on chicken drumsticks and observed a decrease of more than 1 log CFU/g. In addition, our results showed that using some fractions of potato protein in combination with food additives and sodium lactate obtained inactivation levels in excess than 1.66 log CFU/g. The most important C. jejuni counts reductions were always obtained within the first hours after the application of the seasoning combinations onto the chicken meat.
  • Jakava-Viljanen, Miia (Helsingin yliopisto, 2007)
    Lactobacilli, common members of porcine intestinal microbiota, have been considered to be an important group of bacteria in maintaining the stability of gastrointestinal tract (GIT), preventing intestinal infections and supporting intestinal health. Because several species of lactobacilli have GRAS (generally regarded as safe) status and some of them have an ability to interact with intestinal epithelial cells, their possible applications as mucosal vaccine vector and/or probiotics have aroused interest. Selection criteria for lactobacilli to be used as vaccine vector or probiotic include the abilities to adhere to the intestinal epithelium cells and colonize the lumen of the GIT. Bacterial adhesins are often found in hair-like appendages called pili or fimbriae that extend outward from bacterial surface. Alternatively, they can be directly associated with the microbial cell surface. Surface layer proteins (Slps) of lactobacilli have been shown to confer tissue adherence. In this study, S-layer carrying lactobacilli from the intestine and faeces of pigs were isolated and their ability to adhere was studied. Besides the putative binding properties of Slps, a very large number of Slp subunits present in an S-layer make the use of the S-layer structure a very interesting alternative to surface display antigens. Therefore, the aim was to characterize the S-layer proteins. Two new surface layer proteins with potential to be tested as antigen carriers were characterized, and three slp genes were isolated, sequenced, and studied for their expression. To identify the S-layer carrying lactobacilli strains of porcine origin, a polyphasic taxonomic approach was applied. These results indicated that strains from Finland and the related L. sobrius strains, originating from elsewhere, constitute a single species, L. amylovorus, and that the name L. sobrius should be considered as a later synonym of L. amylovorus. The F18 fimbriae carrying Escherichia coli strains cause post-weaning diarrhoea and edema disease in pigs. The adhesin FedF of E. coli F18 fimbriae was characterized. The work aims at developing lactobacilli as a live mucosal vaccine vector for pigs against diseases caused by F18+ E. coli. Oral immunization of weaned piglets with adhesins is known to induce a protective mucosal immune response. Naked FedF appeared to be very unstable but we could produce it as a fusion protein with maltose binding protein (MBP). Specific adhesion to isolated porcine intestinal epithelial cells was demonstrated with MBP-FedF fusions as well as the ability of anti-MBP-FedF antibodies to prevent binding of E. coli F18 to porcine epithelial cells.
  • Kareskoski, Maria (Helsingin yliopisto, 2011)
    Seminal plasma (SP) is the fluid portion of semen, secreted by the epididymides and the accessory glands before and during ejaculation. The stallion s ejaculate is a series of jets that differ in sperm concentration, semen volume and biochemical composition. Before the actual ejaculation, a clear and watery pre-sperm fluid is secreted. The first three jets form the sperm-rich fractions, and contain ¾ of the total number of sperm. The semen volume and sperm concentration in each of the jets decrease towards the end of the ejaculation, and the last jets are sperm-poor fractions with a low sperm concentration. The aims of these studies were to examine the effects of the different SP fractions, and the presence of SP, on sperm survival during storage. Pre-sperm fluid, and semen fractions with a high (sperm-rich) and low (sperm-poor) sperm concentration were collected in five experiments. The levels of selected enzymes, electrolytes and proteins in different SP fractions were determined. These studies also aimed at assessing the individual variation in the levels of the selected SP components and in the effects of SP on spermatozoa. The association between the components of SP and semen quality, sperm longevity, and fertility was examined with a stepwise linear regression analysis. Compared to samples containing SP during storage, centrifugation and the subsequent removal of SP reduced sperm motility parameters during 24 h of cooled storage in all SP fractions, but sperm membrane integrity was not affected. Some of the measured post-thaw motility parameters were also higher in samples containing SP compared to samples stored without SP. In contrast, the proportion of DNA-damaged spermatozoa was greater in the samples stored with SP than those without SP, and this effect was seen in both sperm-rich and sperm-poor fractions. There were no differences in DNA integrity between fractions stored with SP, but the sperm-rich fraction showed less DNA damage than the sperm-poor fraction after SP removal. The differences between fractions in sperm motility after cooled storage were non-significant. The levels of alkaline phosphatase, acid phosphatase and β-glucuronidase were higher in the sperm-rich fractions compared to the sperm-poor fractions, while the concentrations of calcium and magnesium were higher in sperm-poor fractions than in sperm-rich fractions. The concentrations of sodium and chloride were highest in pre-sperm fluid. In the sperm-poor fraction, the level of potassium was associated with the maintenance of sperm motility during storage. The levels of alkaline and acid phosphatase were associated with sperm concentration and the total number of spermatozoa in the ejaculates. None of the measured SP components were correlated to the first cycle pregnancy rate. In summary, the removal of SP improved DNA integrity after cooled storage compared with samples containing SP. There were no differences in the maintenance of sperm motility between the sperm-rich and sperm-poor fractions and whole ejaculates during cooled storage, irrespective of the presence of SP. The lowest rate of DNA damage was found in the sperm-rich fractions stored without SP. In practice, the results presented in this thesis support the use of individual modifications of semen processing techniques for cooled transported semen from subfertile stallions.
  • Leinonen, Merja (Helsingin yliopisto, 2011)
    The primary aim of this thesis was the evaluation of the perfusion of normal organs in cats using contrast-enhanced ultrasound (CEUS), to serve as a reference for later clinical studies. Little is known of the use of CEUS in cats, especially regarding its safety and the effects of anesthesia on the procedure, thus, secondary aims here were to validate the quantitative analyzing method, to investigate the biological effects of CEUS on feline kidneys, and to assess the effect of anesthesia on splenic perfusion in cats undergoing CEUS. -- The studies were conducted on healthy, young, purpose-bred cats. CEUS of the liver, left kidney, spleen, pancreas, small intestine, and mesenteric lymph nodes was performed to characterize the normal perfusion of these organs on ten anesthetized, male cats. To validate the quantification method, the effects of placement and size of the region of interest (ROI) on perfusion parameters were investigated using CEUS: Three separate sets of ROIs were placed in the kidney cortex, varying in location, size, or depth. The biological effects of CEUS on feline kidneys were estimated by measuring urinary enzymatic activities, analyzing urinary specific gravity, pH, protein, creatinine, albumin, and sediment, and measuring plasma urea and creatinine concentrations before and after CEUS. Finally, the impact of anesthesia on contrast enhancement of the spleen was investigated by imaging cats with CEUS first awake and later under anesthesia on separate days. -- Typical perfusion patterns were found for each of the studied organs. The liver had a gradual and more heterogeneous perfusion pattern due to its dual blood flow and close proximity to the diaphragm. An obvious and statistically significant difference emerged in the perfusion between the kidney cortex and medulla. Enhancement in the spleen was very heterogeneous at the beginning of imaging, indicating focal dissimilarities in perfusion. No significant differences emerged in the perfusion parameters between the pancreas, small intestine, and mesenteric lymph nodes. -- The ROI placement and size were found to have an influence on the quantitative measurements of CEUS. Increasing the depth or the size of the ROI decreased the peak intensity value significantly, suggesting that where and how the ROI is placed does matter in quantitative analyses. --- A significant increase occurred in the urinary N-acetyl-β-D-glucosaminidase (NAG) to creatinine ratio after CEUS. No changes were noted in the serum biochemistry profile after CEUS, with the exception of a small decrease in blood urea concentration. The magnitude of the rise in the NAG/creatinine ratio was, however, less than the circadian variation reported earlier in healthy cats. Thus, the changes observed in the laboratory values after CEUS of the left kidney did not indicate any detrimental effects in kidneys. Heterogeneity of the spleen was observed to be less and time of first contrast appearance earlier in nonanesthetized cats than in anesthetized ones, suggesting that anesthesia increases heterogeneity of the feline spleen in CEUS. ---- In conclusion, the results suggest that CEUS can be used also in feline veterinary patients as an additional diagnostics aid. The perfusion patterns found in the imaged organs were typical and similar to those seen earlier in other species, with the exception of the heterogeneous perfusion pattern in the cat spleen. Differences in the perfusion between organs corresponded with physiology. Based on the results, estimation of focal perfusion defects of the spleen in cats should be performed with caution and after the disappearance of the initial heterogeneity, especially in anesthetized or sedated cats. Finally, these results indicate that CEUS can be used safely to analyze kidney perfusion also in cats. Future clinical studies are needed to evaluate the full potential of CEUS in feline medicine as a tool for diagnosing lesions in various organ systems.
  • Nuotio, Lasse (Helsingin yliopisto, 2006)
    The monitoring and control of infectious animal diseases, limiting or prevention of their spread and efforts towards their eradication are central tasks of the veterinary civil service. In addition to the cost-effectiveness of prophylaxis over disease and treatment, the animal welfare aspect is also involved. The purpose of this work is to review, describe and assess the available control measures against selected viral infections or diseases of domestic ruminants. The selected infections or diseases are bovine viral diarrhoea / mucosal disease (BVD), infectious bovine rhinotracheitis / infectious pustular vulvovaginitis (IBR), enzootic bovine leukosis (EBL) and maedi-visna (MV) of sheep. Decisive control and eradication measures are necessarily based on the biological, veterinary and diagnostic characteristics of the afflictions, as well as on their epidemiology in terms of the intrinsic determinants of the hosts, host-agent relationships, transmission of the infection and occurrence of these infections or diseases. This information is compiled with special reference to available or possible control and eradication measures, and those employed in major cattle and sheep producing countries are outlined and assessed briefly. The domestic and EU legislation that forms the official framework for disease control and eradication, as well as the official measures and the voluntary programmes are reviewed. The first recorded entries of the occurrence of BVD and EBL in Finland date back to the 1960s, those of IBR to the beginning of the 1970s and of MV to the beginning of the 1980s. Large-scale surveillance and health monitoring among dairy, suckler-cow and beef herds and sheep flocks, starting during the first half of the 1990s, enabled the estimation of actual prevalences of these infections and diseases. The successful eradication of IBR and EBL in 1994 and 1996, respectively, and the significant reduction in the occurrences of BVD and MV from 1990 to date, are reported in detail. The efficacies of the official control and eradication measures and of the voluntary control programmes are analyzed further with a heuristic formulation for the infection reproduction number (R), i.e. the number of secondary cases produced by one infective animal. The influence of the measures is resolved into probability of transmission, frequency of infectious contacts and length of the infectious period, and the impact of the measures on each component is graded on a three-step scale. The conclusion is drawn that the official measures complemented by voluntary actions for control and eradication have for the most part been adequate. In the case of BVD the decisive measures for final eradication have only been available since 2004 and their impact will be seen in the next few years. The role of continued surveillance and health monitoring for both overseeing the situation with BVD and MV, and maintaining an IBR and EBL-free status is emphasized.
  • Hyytiäinen, Heli (Helsingin yliopisto, 2015)
    Stifle dysfunction is one of the most common reasons for canine hindlimb lameness and an indication for dogs referral to physiotherapy. Until now, there has been a lack of testing batteries in animal physiotherapy, although these are an important part of the evaluation process in various patient groups in human physiotherapy. Using 64 dogs, 43 with stifle dysfunction and 21 healthy dogs, congruity between fourteen physiotherapeutic evaluation methods, commonly used in dogs with stifle dysfunction, and six evaluation methods used by a veterinarian was evaluated. The eight best methods were chosen as items constituting a testing battery, the Finnish Canine Stifle Index (FCSI). The numerical scale of the testing battery was 0-263. Cronbach s alpha for the internal reliability of the total FCSI score was good (0.727). Two cut-offs for the total score were set: 60 and 120, separating adequate, compromised and severely compromised performance level, based on their high sensitivities and specificities. Another 57 dogs, 29 with some type of stifle dysfunction, 17 with some musculoskeletal disease other than stifle dysfunction and 11 healthy dogs, were used to further study the psychometric properties of the testing battery. The dogs with stifle dysfunction showed a significant (P < 0.001) decrease in FCSI total score (93.3 ± 62) compared with the two other groups (29.5 ± 39.6 and 11.7 ± 21.0), demonstrating good responsiveness of the FCSI. Also the inter-tester reliability was excellent (ICC 0.784), with no significant differences between three physiotherapists performing the FCSI. In conclusion, the overall functionality and outcome of rehabilitation in dogs with stifle dysfunction can be reliably evaluated with the new testing battery, the FCSI, developed here.
  • Õkva, Kai (Helsingin yliopisto, 2012)
    Discussions on laboratory animal welfare issues often refer to the Three Rs replacement, reduction and refinement. Replacement means substituting living animals with non-sentient systems; reduction refers to using fewer animals and refinement causing less pain, suffering and distress to the animals or improving their welfare. This work is focusing on two R-s: reduction and refinement in mice. If one considers reduction as meaning obtaining trustworthy information from using fewer animals, then this can be achieved by improved research strategies, better experimental design and more sophisticated statistical analyses. One can reduce variation within the group by using isogenic inbred animals or by finding ways to reduce variation in outbred animals. One approach to achieve reduction in outbred animals is to include litter and individual features of the animals, e.g weight dynamics, in the statistical analyses. Since the elevated plus-maze (EPM) test is one of the most common tests to evaluate anxiety-like behaviour, it was used to assess the possible effects of litter and weight on the behaviour of outbred mice and the effects of environmental enrichment (EE) on the behaviour of inbred mice. As a research tool, the effects of acute or chronic administration of ethanol or acute therapy with the nitric oxide synthase (NOS) inhibitor L-NOARG were examined in outbred NIH/S mice. The administration of L-NOARG had no effect on the behaviour of mice after acute or chronic ethanol administration but attenuated the anxiogenic effect of ethanol withdrawal. The litter from which the mice had originated had a significant effect on their behaviour in the EPM test. The behavioural indices of mice, originating from different litters, tended to be above or below the mean of the corresponding drug-treatment group, irrespective of the drug treatment. Litter had a significant effect on the initial weight and also on the weight changes occurring during the adaptation period and ethanol inhalation. An approach to refinement, EE, has been introduced to create more natural species-specific living conditions for laboratory animals. At the same time, it has been claimed that EE can affect the results of behavioural studies and also increase variation. The effects of different types of EE and different time periods were studied in inbred C57BL/6J and BALB/c mice. The exposure of male C57BL/6J mice to the different types of EE objects in the form of cage furniture (CF) -nest box, corner and stairs- induced an anxiolytic-like effect in the EPM test and tended to increase the locomotor activity of mice. This apparent anxiolytic-like effect was most pronounced in the third week. The CF, in the form of modified Tapvei OY mouse stairs, produced an anxiolytic-like effect and increased the locomotor activity in female C57BL/6J mice, but not in BALB/c female mice. In conclusion, the NOS inhibitors may have effects on the behavioural changes caused by ethanol withdrawal. Information about the litter of outbred mice could and should be used in statistical analysis in order to reduce variation and the number of mice needed. In the EPM test, different CF items induced an anxiolytic-like effect in male and female C57BL/6J mice, but not in BALB/c female mice. This effect depended on the type of objects and was influenced by time. This anxiolytic like effect can be interpreted as refinement of the housing by improving animal welfare. The effects of CF should be considered in planning enrichment programs for housing institutions, in designing behavioural experiments and in analyzing the results obtained. Therefore the evaluation of CF could provide the valuable information and it is recommended that CF manufacturers collate and distribute the refinement results on the specific CF items they produce.
  • Autio, Karoliina (Helsingin yliopisto, 2015)
    Cancer is one of the most common reasons for death in dogs, cats and humans. New therapeutic modalities are necessary to improve disease outcome. One promising approach is oncolytic virotherapy. Until now, the only oncolytic virus evaluated in a clinical trial in veterinary medicine has been canine oncolytic adenovirus, but a clinical trial has been started with oncolytic vaccinia virus (VV) in pet dogs. In cats, oncolytic viruses have not been evaluated in clinical settings. Tumour treatment in dogs and cats could also serve as a model for human cancer therapy. The purpose of the thesis was to evaluate preclinically whether genetically modified oncolytic VV and Semliki Forest virus (SFV) could offer a new treatment modality for dogs and cats. Oncolytic VV was rendered tumour selective by a dual ablation of vaccinia growth factor and thymidine kinase, making it more cancer-specific than previously used VVs in veterinary research. To further increase the efficacy of the VV, an immunostimulatory gene, CD40L, was added to the virus backbone. Avirulent SFV A7(74) has a natural trophism for cancer cells and was not genetically manipulated. Both viruses infected and killed tested cancer cell lines, and VV also infected most of the primary surgical tumour tissues tested. In the nude mouse xenograft model, double deleted VV (vvdd) significantly reduced tumour growth. Interestingly, when in intact monolayers, SCCF1 cells were not killed by VV, but secreted infectious, morphologically abnormal virions. One dog experienced a possible seizure after VV administration, but no other serious adverse events occurred. Vaccinia DNA declined quickly in the blood after virus administration, but was still detectable one week later by qPCR. Only samples taken directly after the VV infusion contained infectious virus, which was not found in any other blood, saliva, urine or faecal samples. Necropsies did not reveal any pathological changes associated with virus administrations. In conclusion, our results show that oncolytic VV and SFV can infect and kill tested canine cancer cell lines, and that VV has antitumoral activity in the mouse xenograft model and it can infect fresh tumour biopsies. In addition, intravenous administration of the viruses did not induce life-threatening adverse events in healthy dogs. These agents thus warrant further evaluation in veterinary medicine.
  • Rintakoski, Simo (Helsingin yliopisto, 2014)
    In Denmark, Finland, Norway and Sweden the National Dairy Disease Registers (NDDRs) collect and store disease information at the individual cow level. Because these registers are monitored nationally they offer access to data that cover most of the dairy population in each country. Data from these registers are used, for example, for herd health and animal welfare assessments, production management, genetic evaluations and epidemiologic research. Since the register data are not usually designed for research purposes their quality is unknown. Understanding the magnitude of data loss during data transfer and human influence in disease recording process is important. The knowledge will enhance the ability to improve register quality, which in turn will improve reliability of frequency measure calculations from the register data. This thesis investigated the quality (measured as completeness and correctness) of the Finnish NDDR and compared register qualities among the four countries. In Finland the quality of recorded information was excellent, but approximately 17% of disease information was lost during the data transfer steps. A large proportion of the data loss was due to artificial insemination (AI) technicians not transferring events. The majority of those events occurred close to culling of the cow, suggesting early removal of the cow health cards so that AI technician did not have access to those cards. Additionally, fewer diagnostic events were transferred from purchased cows compared to those born on farm suggesting problems with animal identification. An efficient way to improve completeness in the Finnish NDDR is to have veterinarians electronically transfer diagnostic information during farm visits. The Finnish system has already been modified accordingly. This thesis also showed how register quality for four reproductive disorders (metritis, retained placenta, assisted calving and oestrous disturbances) varied among the four countries. Metritis and oestrous disturbance events were well represented in the NDDRs. Farmer-observed completeness (the proportion of all farmer observations that were recorded in the NDDR) was around 0.80 and did not differ significantly among the countries. Assisted calving and retained placenta events showed more among-country variation. Farmer-observed completeness was highest in Denmark and lowest in Finland, ranging between 0.31 and 0.89. The comparison of completeness-adjusted incidences to incidences calculated from the registers showed that incidences were underestimated for assisted calving and retained placenta. Underestimation was highest in Finland. This thesis also demonstrated how both farmer and veterinary intentions toward starting veterinary treatment of mild clinical mastitis could explain the reasons for different mastitis incidence rates among the countries. The results suggest that when intentions towards veterinary treatment are greater, mild cases receive veterinary treatment more often than when intentions are reduced. Therefore, greater farmer and veterinarian intentions can increase the disease incidence calculated from the NDDR data. According to the study, intentions were greater in Denmark than in other countries.
  • Markkula, Annukka (Helsingin yliopisto, 2013)
    Listeria monocytogenes causes potentially fatal illness to susceptible people and is found in various foods. It typically enters processed foods via a contaminated processing environment, in which it may have persisted for years. To study the role of raw material as a potential source of contamination of food processing plants by L. monocytogenes, the prevalence and genetic diversity of this species in tonsils of pigs and in raw fish was examined. A total of 14% and 4% of tonsils of pigs and raw fish, respectively, harboured L. monocytogenes. From 38 pig tonsil isolates and 11 raw fish isolates, 24 and nine different types were recovered using pulsed-field-gel electrophoresis (PFGE) typing. The results indicate that a wide variety of L. monocytogenes strains enters pork slaughterhouses and fish processing plants in the raw materials, which are thus potential sources of direct or indirect contamination of processing plants by this pathogen. Since identical PFGE types were recovered from both raw and processed fish, it is likely that raw fish are an initial source of the L. monocytogenes found in processed fish. Some strains entering a plant along with raw fish may contaminate and persist in the processing environment, causing recurrent contamination of the final products via contact surfaces. Alternatively, L. monocytogenes strains in raw fish may survive non-listericidal processes, resulting in contamination of the final product. To identify novel factors contributing to survival of L. monocytogenes in food processing environment, the roles of specific genes in stress response were investigated, using flhA and motA that encode flagellar factors involved in cold stress tolerance, and lmo0866, lmo1246, lmo1450, and lmo1722 encoding DEAD-box RNA helicases involved in cold, heat acid, alkali, osmotic, ethanol, and oxidative stress tolerance. Increased relative transcription levels of flhA, motA, lmo0866, lmo1450, and lmo1722, restricted growth of the single gene deletion mutant strains EGD-eΔflhA, EGD-eΔmotA, Δlmo0866, Δlmo1450, and Δlmo1722 at 3°C, and increased minimum growth temperatures of Δlmo0866, Δlmo1450, and Δlmo1722 revealed that FlhA, MotA, Lmo0866, Lmo1450, and Lmo1722 had roles in growth of L. monocytogenes EGD-e under cold stress conditions. The restricted growth of Δlmo0866 in 3.5% ethanol, and its increased maximum growth temperature and growth rate at 42.5°C, indicated that Lmo0866 had roles also in ethanol and heat stress tolerance of strain EGD-e. The role of Lmo1450 in the growth of strain EGD-e under heat, alkali, and oxidative stress conditions was shown by the restricted growth rate of Δlmo1450 at 42.5°C, in pH 9.4, and in 5 mM H2O2. The slightly decreased growth rate and maximum optical density of Δlmo1246 at 3°C indicated that the role of Lmo1246 in cold stress tolerance was negligible. Under all the other conditions, the growth of Δlmo1246 and the wild-type EGD-e were identical, suggesting that Lmo1246 had no role in growth of L. monocytogenes EGD-e under heat, pH osmotic, ethanol, or oxidative stress conditions. The deletion of flhA, motA, lmo0866, lmo1450, and lmo1722 impaired the motility of strain EGD-e, whereas the motility of Δlmo1246 did not differ from that of the wild type. This indicates that DEAD-box RNA helicases Lmo0866, Lmo1450, and Lmo1722 have roles in motility of strain EGD-e. Moreover, these results suggest that motility and cold stress tolerance of L. monocytogenes are linked, and that motile flagella may be needed for full cold stress tolerance of strain EGD-e.