Murine colonization model by Campylobacter coli DSPV458.

AIMS: To generate a murine experimental model of colonization by Campylobacter coli DSPV458. METHODS AND RESULTS: Twelve adult Balb/cCmedc female mice were housed in a treated group (T-G) and a control group (C-G) for 4 weeks. Both experimental groups received antibiotics for 5 days during the first week. The T-G was administered with 6.68log10 CFU of C. coli DSPV458 by oesophageal gavage. Necropsies were performed weekly to evaluate translocation and intestinal colonization in the spleen and liver and in the ileum and cecum respectively. Samples were cultured to quantify intestinal microbiota members. Faeces were cultured weekly for a C. coli DSPV458 count. Campylobacter coli DSPV458 was isolated from all the inoculated mice. The recovered level of C. coli DSPV458 was, on average, 6.9 log10 CFUg-1 , 8.0 log10 CFUg-1 and 1.6 log10 CFUg-1 in faeces, cecum and ileum respectively. Colonization by C. coli DSPV458 does not alter the normal clinical and physiological status. CONCLUSIONS: Campylobacter coli DSPV458 does not have an invasive capacity, and the model is suitable for evaluating strategies to reduce intestinal loads. SIGNIFICANCE AND IMPACT OF STUDY: Farm animals have an important impact on thermotolerant Campylobacter transmission to humans. Extremely few colonization models by C. coli have been reported to date. In food-producing animals, infection is mild or absent and thermotolerant Campylobacter colonize the intestines of animals. Colonization models are specific models that do not cause infection as they do not generally result in diarrhoea or other signs of disease. Therefore, this model will allow to evaluate the evolution of colonization by thermotolerant Campylobacter and the alternative tools development to antibiotics that limit their colonization in food-producing animals.

Effect of probiotic Lactiplantibacillus plantarum and chestnut flour (Castanea sativa mill) on microbiological and physicochemical characteristics of dry-cured sausages during storage.

The effect of chestnut flour (Castanea sativa Mill) on L. plantarum viability and physicochemical characteristics in a dry-cured sausage (Longaniza de Pascua) during storage is discussed. Four batches were prepared: CL with 3% chestnut flour added; CPL with 3% chestnut flour and 8.5 log CFU/g L. plantarum added; PL with 8.5 log CFU/g L. plantarum added and L, the batch control. The sausages were stored at 4 °C and 20 °C, and vacuum packed for 43 d. L. plantarum viability was affected by storage time (P < 0.001). However, higher L. plantarum counts at the final of storage were reached due to chestnut flour addition (P < 0.001). At room storage, chestnut flour caused a higher increase in TBARS values (P = 0.022). Nevertheless, all lipid oxidation treatments were in the range of accepted values at the sensory detection level. In conclusion, Longaniza de Pascua can be kept at 4 °C or 20 °C for 43 d without causing any rancidity problems.

Protective effect of Lactiplantibacillus plantarum LP5 in a murine model of colonisation by Campylobacter coli DSPV458.

Thermotolerant Campylobacter species are the leading cause of foodborne bacterial diarrheal disease worldwide. Campylobacter coli, abundant in pigs and pork products, have been identified as a source of human infection. In this study, we propose the use of Lactiplantibacillus plantarum LP5 as a probiotic to reduce colonisation of this intestinal pathogen in a murine colonisation model of C. coli DSPV458. Six-week-old adult female Balb/cCmedc mice were housed in groups: Control, Campy and Pro-Campy. Control and Pro-Campy groups received antibiotics for 5 days and the Campy group for 12 days. Pro-Campy group was inoculated for 7 days with 8.78 log10 cfu total of L. plantarum LP5 suspended in De Man, Rogosa and Sharpe broth. All groups were inoculated with 6.72 log10 cfu of C. coli DSPV458 suspended in brain heart infusion broth. L. plantarum LP5 was recovered only in the Pro- Campy group. C. coli DSPV458 was recovered at higher levels in the Control and Campy groups. The differences with the Pro-Campy group were significant. As regards faeces, Control and Campy groups reached 7.41 and 7.84 log10 cfu/g, respectively, and the Pro-Campy group only 4.62 log10 cfu/g. In the caecum, Control and Campy groups reached 8.01 and 9.26 log10cfu/g, respectively, and the Pro-Campy group only 4.51 log10 cfu/g. In the ileum, Control and Campy groups reached 3.43 and 3.26 log10 cfu/g, respectively, and the Pro-Campy group did not show detectable levels. The reduction of C. coli DSPV458 in the Pro-Campy group compared to the Control group in faeces, caecum and ileum was 99.55, 99.98 and 100%, respectively. Animals were maintained under normal health conditions, and haematological parameters were within the standard values for Balb/cCmedc. The incorporation of a probiotic generated a protective effect in the mice colonisation model. The protective effect would also apply to intestinal colonisation by indigenous enterobacteria. Therefore, the strategy used in this study is of great importance to understand the protection mechanisms in a murine model, as well as its application in food-producing animals.

In vitro screening and in vivo colonization pilot model of Lactobacillus plantarum LP5 and Campylobacter coli DSPV 458 in mice.

The objective of this work was to determine the antibacterial effect of Lactobacillus plantarum strains of pork origin against Campylobacter coli strains, and to conduct experimental colonization pilot models in mice for both microorganisms. Inhibition assays allowed evaluation and selection of L. plantarum LP5 as the strain with the highest antagonistic activity against C. coli and with the best potential to be used in in vivo study. Adult 6-week-old female Balb/cCmedc mice were lodged in two groups. The treated group was administered with 9.4 log10CFU/2 times/wk of L. plantarum LP5. L. plantarum LP5 was recovered from the feces and cecum of the inoculated mice. However, when bacteria stopped being administered, probiotic counts decreased. Experimental colonization with C. coli was carried out in five groups of mice. All animals were treated with antibiotics in their drinking water to weaken the indigenous microbiota and to allow colonization of C. coli. Four groups were administered once with different C. coli strains (DSPV458: 8.49 log10CFU; DSPV567: 8.09 log10CFU; DSPV570: 8.46 log10CFU; DSPV541: 8.86 log10CFU, respectively). After 8 h, mice inoculated with different C. coli strains were colonized because the pathogen was detected in their feces. L. plantarum LP5 tolerated the gastrointestinal conditions of murine model without generating adverse effects on the animals. C. coli DSPV458 colonized the mice without causing infection by lodging in their digestive tract, thus generating a reproducible colonization model. Both models combined could be used as protection murine models against pathogens to test alternative control tools to antibiotics.

Possible reservoirs of thermotolerant Campylobacter at the farm between rearing periods and after the use of enrofloxacin as a therapeutic treatment.

Campylobacteriosis is a zoonosis and the most frequent cause of food-borne bacterial enteritis in humans. C. jejuni and C. coli are the most common species implicated in campylobacteriosis. Broilers and their products are considered the most important food sources of human infections. The aim of the present study was to evaluate the presence of thermotolerant Campylobacter in different reservoirs at the farm, and the permanence of this pathogen during four consecutive rearing periods. The samples were taken from the same house farm in the downtime period and during the last week of broiler rearing, prior to their slaughter during four consecutive cycles. Different reservoirs as potential sources of Campylobacter were analysed. The prevalence of Campylobacter in vectors was 23% in A. diaperinus larvae, 20% in wild birds, 13% in A. diaperinus adults, and 9% in flies; as regards fomites, the prevalence was 50% in workers' boots, 27% in litter, and 21% in feed, while in broilers it was 80%. Campylobacter jejuni was the most detected species (51%) in the samples analysed. In addition, some Campylobacter genotypes persisted in the house farm throughout consecutive rearing periods, indicating that those strains remain during downtime periods. However, our study could not identify the Campylobacter sources in the downtime periods because all the samples were negative for Campylobacter isolation. In addition, a remarkable finding was the effect of the use of enrofloxacin (as a necessary clinical intervention for flock health) in cycle 3 on the Campylobacter population. No Campylobacter could be isolated after that clinic treatment. Afterwards, we found a greater proportion of C. coli isolates, and the genotypes of those isolates were different from the genotypes found in the previous rearing periods. In conclusion, the effect of the use of enrofloxacin during the rearing period changed the Campylobacter species proportion, and this finding is particularly interesting for further evaluation. Furthermore, more studies should be conducted with the aim of detecting the Campylobacter sources between rearing periods.

Molecular epidemiology of Campylobacter jejuni isolates from the broiler production chain: first report of MLST profiles in Argentina.

Campylobacter jejuni is an important foodborne pathogen with global distribution. We describe a genotyping study of a collection of C. jejuni (n=137) isolated from different broiler farms and from multiple sites along the processing line in a slaughterhouse in Argentina during 2011, 2012 and 2015. The isolates were genotyped using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Based on the PFGE results, the isolates were grouped into 26 pulsotypes. Subsequently, the isolates representing these 26 pulsotypes were chosen for MLST genotyping, which identified 16 different sequence types (STs) and 6 clonal complexes (CCs) (21, 45, 48, 353, 354, 446). Several of the STs (n=7) have not been previously reported in the PubMLST.org database. The most prevalent CCs were 21, 45 (both associated with human campylobacteriosis worldwide) and 353. This study showed high genetic diversity among C. jejuni in the broiler production environment in Argentina with novel MLST genotypes.

Worldwide meta-analysis of the prevalence of Campylobacter in animal food products.

The objective of this meta-analysis was to summarize available information on the prevalence of thermotolerant Campylobacter in different animal food products. A number of multilevel random-effect meta-analysis models were fitted to estimate mean prevalence of thermotolerant Campylobacter and to compare them among animal food products (cattle, pigs, broiler, hen, goat, sheep). The mean prevalence of Campylobacter spp. in animal food products was 29.6% (95% CI 27.6%-31%), and the mean prevalence of C. jejuni and C. coli were 19.3% and 9.7%, respectively. The prevalence of Campylobacter spp. was higher in products whose sources were broiler meat (p-estimate = 47.8%; 95% CI 44.9%-50.6%). C. jejuni was mainly observed in broiler meat where prevalence estimate (p-estimate) was 33.7% (95% CI 30.7%-36.8%). On the other hand, C. coli was observed in broiler meat (p-estimate = 14.1%; 95% CI 12.3%-16.1%) and sheep meat (p-estimate = 11.0%; 95% CI 3.6%-29.1%). The animal food products with the lowest prevalence of Campylobacter spp. were milk and dairy products (p-estimate = 3.5%; 95% CI 1.8%-6.5%), eggs (p-estimate = 4.0%; 95% CI 1.4%-10.7%), sausage (p-estimate = 9.4%; 95% CI 3.3%-24.0%), This meta-analysis concluding that C. jejuni is the most prevalent species worldwide and broiler meat is the main contamination source for human. The prevalence of Campylobacter species has public health importance and national authorities must monitor the situation in each country with the aim to establish the appropriate risk management measures.

Impact of lyophilized Lactobacillus salivarius DSPV 001P administration on growth performance, microbial translocation, and gastrointestinal microbiota of broilers reared under low ambient temperature.

This study was undertaken with the aim of investigating the effects of dietary supplementation of probiotic strain Lactobacillus salivarius DSPV 001P on growth performance, microbial translocation, and gastrointestinal microbiota of broilers reared under low ambient temperature. Two hundred and forty, one-day-old male Cobb broilers were randomly distributed into two treatment groups, a probiotic group and a control group, with four replicates per treatment and 30 broilers per replicate. The temperature of the broiler house was maintained at 18-22°C during the first three weeks, after which the temperature was at range of 8°C to 12°C. The results showed that probiotic treatment significantly improved body weight of broilers when compared with the control group. After 42days, the weight means were 2905±365.4g and 2724±427.0g, respectively. Although there were no significant differences, dietary inclusion of L. salivarius tended to increase feed intake and to reduce feed conversion ratio during the six-week experimental period. Similarly, supplementation tended to reduce the rate of mortality, with 12 deaths occurring in the probiotic group, and 20 in the control group. However, no differences were observed in intestinal bacterial concentrations of Enterobacteriaceae, E.coli, and lactic acid bacteria in both crop and caecum among treatments. Through our study, it appears that L. salivarius DSPV 001P was non-pathogenic, safe and beneficial to broilers, which implies that it could be a promising feed additive, thus enhancing the growth performance of broilers and improving their health.

Quantification of FITC-labelled probiotic Lactobacillus salivarius DSPV 001P during gastrointestinal transit in broilers.

The knowledge related to the fate of probiotics in the complex environment of the intestinal microbiota in broilers is just beginning to be elucidated; however, it is not yet well understood. A good method to investigate the mechanisms by which probiotics mediate their effects is to mark probiotic bacteria and trace them. The aim of this research was to develop a new method to estimate in vivo fluorescein isothiocyanate (FITC)-labelled Lactobacillus salivarius DSPV 001P counts during passage through the gastrointestinal tract (GIT) of broilers. Forty-five, 1 d old Cobb broilers were used in this trial. Programmed necropsies were performed 30 min, 6 h, and 12 h after the administration of the probiotic bacterium, and samples of liver, crop, duodenum, caecum, and bursa of fabricius were collected. To determine the spatial and temporal transit of L. salivarius DSPV 001P in broilers, the number of bacteria as well as its respective fluorescent signal produced by FITC were measured. In order to observe the relationship between the variables, a logistic regression analysis was applied. The amount of fluorescence could be used as an indicator of fluorescent probiotic bacteria in the crop and duodenum 30 min after probiotic bacterium supplementation. In addition, the fluorescent signal could be used to estimate bacterial counts in caecum 6 and 12 h after L. salivarius DSPV 001P administration. To the best of our knowledge, this research is the first in vivo trial to employ the bacterial FITC-labelling technique in order to enumerate probiotic bacteria during gastrointestinal transit in broilers.

Poultry blood from slaughterhouses: development of a biopreservation system to improve microbiological quality prior to transforming blood into by-products.

The aim of this study was to investigate the use of indigenous lactic acid bacteria (LAB) with specific additives as a Biopreservation System (BS) for poultry blood during its storage in slaughterhouses. The BS consisted of two LAB (Enterococcus faecalis DSPV 008SA or Lactobacillus salivarius DSPV 032SA) with 4 additives (lactose 2 g/l, yeast extract 0.4 g/l, ammonium citrate 0.4 g/l and NaCl 1 g/l). After 24 h storage at 30ºC, lower counts of enterobacteria, coliforms, Pseudomonas spp. and Staphylococcus aureus were evident in blood treated with the BS than in untreated blood. The ability of LAB to prevent haemolysis was evident. A decrease in pH was associated with control of spoilage microorganisms but it needed to be regulated to prevent coagulation of proteins. On the basis of these results it is recommended to supplement blood with a BS to avoid undesirable changes during blood storage before processing.

Probiotic effect on calves infected with Salmonella Dublin: haematological parameters and serum biochemical profile.

The aim of this study was to evaluate the effect of a probiotic/lactose inoculum on haematological and immunological parameters and renal and hepatic biochemical profiles before and during a Salmonella Dublin DSPV 595T challenge in young calves. Twenty eight calves, divided into a control and probiotic group were used. The probiotic group was supplemented with 100 g lactose/calf/d and 1010 cfu/calf/d of each strain of a probiotic inoculum composed of Lactobacillus casei DSPV318T, Lactobacillus salivarius DSPV315T and Pediococcus acidilactici DSPV006T throughout the experiment. The pathogen was administered on day 11 of the experiment, at an oral dose of 109 cfu/animal (LD50). Aspartate aminotransferase (AST), gamma glutamyl transpeptidase (GGT), urea, red blood cells, haemoglobin, haematocrit, mean cell haemoglobin (MCH), mean corpuscular volume, mean corpuscular haemoglobin concentration (MCHC), white blood cells, lymphocytes, neutrophils, band neutrophils, monocytes, eosinophils, basophils and the neutrophils/lymphocytes ratio were measured on days 1, 10, 20 and 27 of the experiment. In addition, animals were necropsied to evaluate immunoglobulin A (IgA) production in the jejunal mucosa. The most significant differences caused by the administration of the inoculum/lactose were found during the acute phase of Salmonella challenge (9 days after challenge), when a difference between groups in neutrophils/lymphocytes ratio were detected. These results suggest that the probiotic/lactose inoculum administration increases the calf's ability to respond to the disease increasing the systemic immune response specific. No differences were found in haemoglobin, haematocrit, MCH, MCHC, AST, urea, GGT, band neutrophils, eosinophils, monocytes and IgA in the jejunum between the two groups of calves under the experimental conditions of this study. Further studies must be conducted to evaluate different probiotic/pathogens doses and different sampling times, to achieve a greater understanding of the effects of this inoculum on intestinal infections in young calves and of its mechanism of action.

Quantitative Risk Assessment of Human Trichinellosis Caused by Consumption of Pork Meat Sausages in Argentina.

In Argentina, there are three known species of genus Trichinella; however, Trichinella spiralis is most commonly associated with domestic pigs and it is recognized as the main cause of human trichinellosis by the consumption of products made with raw or insufficiently cooked pork meat. In some areas of Argentina, this disease is endemic and it is thus necessary to develop a more effective programme of prevention and control. Here, we developed a quantitative risk assessment of human trichinellosis following pork meat sausage consumption, which may be used to identify the stages with greater impact on the probability of acquiring the disease. The quantitative model was designed to describe the conditions in which the meat is produced, processed, transported, stored, sold and consumed in Argentina. The model predicted a risk of human trichinellosis of 4.88 × 10(-6) and an estimated annual number of trichinellosis cases of 109. The risk of human trichinellosis was sensitive to the number of Trichinella larvae that effectively survived the storage period (r = 0.89), the average probability of infection (PPinf ) (r = 0.44) and the storage time (Storage) (r = 0.08). This model allowed assessing the impact of different factors influencing the risk of acquiring trichinellosis. The model may thus help to select possible strategies to reduce the risk in the chain of by-products of pork production.

Probiotics and broiler growth performance: a meta-analysis of randomised controlled trials.

1. The aim of this meta-analysis was to investigate the effects of probiotics on the growth performance of broilers. PubMed, Scopus and Scholar Google databases were searched in all languages from 1980 to 2012. The studies in the meta-analysis were only selected if they were randomised and controlled experiments using broilers without apparent disease and the results were published in peer-reviewed journals. 2. A total of 48 and 46 studies were included to assess probiotic effects on body weight gain (BWG) and feed efficiency (FE), respectively. Probiotics increased BWG compared to controls (SMD = 0.661, 95% CI 0.499 to 0.822) and improved FE (SMD = - 0.281, 95% CI -0.404 to -0.157) in the pooled standardised mean difference random effect model, considering the source of heterogeneity and publication biases. However, there are evidences of publication bias and heterogeneity, so the results of this meta-analysis should be considered with caution. Applying the Duval and Tweedie's trim-and-fill methods, the adjusted value for BWG was 0.0594 (95% CI -0.122 to 0.242), and the adjusted value for FE did not show any modifications. 3. The meta-analysis showed that application of probiotics via water resulted in greater BGW and FE than administration through the feed. The effect was not related to the use of mono-strain or multi-strain probiotics, although it may depend on the strain used. The number of broilers and the duration of the experiments had an impact on the outcomes. 4. Additional studies should be conducted with the aim to identify the covariates which can explain the differences in the estimated effect sizes.

Occurrence of thermotolerant Campylobacter spp. at different stages of the poultry meat supply chain in Argentina.

AIMS: The objectives of this study were to investigate the occurrence and concentration of thermotolerant Campylobacter spp. at different stages of the poultry meat supply chain in Argentina. METHODS: Three integrated poultry companies were sampled. Each supply chain was considered at different stages from the reproductive farm to chicken meat at a retail market. The stages sampled were: (a) hens from breeder flocks, (b) eggs in the incubator, (c) broiler chickens in flocks (aged <1 week and >5 weeks), (d) chickens at a slaughterhouse, and (e) chicken meat at a retail market. The chickens sampled along each supply chain were in the same batch. Samples collected were: (a) cloacal samples from hens and chickens on the farms, (b) fertile eggs, (c) feed, water and litter from flocks, (d) chicken carcasses from the slaughterhouse and retail market, and (e) caeca and livers from the slaughterhouse. Samples obtained were examined for Campylobacter spp. The isolates were biotyped and the genus and species identified by PCR. Campylobacter spp. on chicken carcasses at slaughterhouse and retail market were enumerated. RESULTS: The highest proportions of Campylobacter positive samples were observed in carcasses at retail (25/30, 83.3%) and faecal samples from breeding hens (27/45, 60.0%). Only 3.3% (3/90) samples collected from broiler chickens aged <1 week were positive, but the percentage of positive samples had risen to 28.9% (26/90) by the end of the rearing period. The proportions of Campylobacter positive carcasses and caecal contents at the slaughterhouse were both 33.3% (10 of 30 samples each). The concentration of Campylobacter contamination observed on carcasses at retail markets ranged from no bacteria/carcass to 3.71 log10 cfu/carcass. CONCLUSIONS: The data obtained provide essential information for future quantitative risk assessments aiming to estimate the probability of a person contracting campylobacteriosis following consumption of broiler meat in Argentina. CLINICAL RELEVANCE: The proportions of Campylobacter-positive samples found in this preliminary study indicate that a large proportion of the cases of human gastroenteritis in Argentina may be due to this pathogen. Human cases of gastroenteritis should be studied in greater detail and measures should be developed to reduce the proportion of poultry products that are contaminated by Campylobacter species.

Quantitative risk assessment of human campylobacteriosis by consumption of salad cross-contaminated with thermophilic Campylobacter spp. from broiler meat in Argentina.

Here, we developed a quantitative risk assessment for thermophilic Campylobacter spp. related to the consumption of salad prepared alongside broiler meat. The assessment considered initial contamination levels, cross-contamination and decontamination events during the broiler slaughter process and distribution, and storage and consumption patterns in Argentina and other Latin American countries. The model predicted an infection risk of 3.32×10(-4) per serving. This estimation was variable according to the dose-response model used. Considering the number of chickens slaughtered annually in Argentina, the estimated number of people who could suffer campylobacteriosis related to poultry meat consumption was, on average, 484,304. The risk of human campylobacteriosis was most sensitive to the probability of infection from a Campylobacter (r=0.72), the number of Campylobacter spp. per serving (r=0.40), the frequency of washing the cutting board (r=-0.31), the preparation of raw poultry before salad using the same cutting board (r=0.14), and the frequency of hand washing (r=-0.14). The most sensitive stages of the process identified through the risk assessment can be used as a basis for measures of risk management. Public campaigns on hygiene habits during food preparation at home should focus on the importance of washing the cutting board before preparing raw and ready-to-eat foods and of washing the hands during food preparation.

Impact of probiotic administration on the health and fecal microbiota of young calves: a meta-analysis of randomized controlled trials of lactic acid bacteria.

Before weaning, dairy calves are susceptible to many pathogens which can affect their subsequent performance. The use of lactic acid bacteria (LAB) has been identified as a tool to maintain the intestinal microbial balance and to prevent the establishment of opportunistic pathogenic bacterial populations. However, a consensus has not been reached as to whether probiotics may be effective in reducing the prevalence of gastrointestinal diseases in young calves. The aim of this meta-analysis was to assess the effect of probiotics on diarrhea incidence and the intestinal microbial balance. LAB supplementation has been shown to exert a protective effect and to reduce the incidence of diarrhea (relative risk, RR=0.437, 95% confidence interval (CI) 0.251-0.761). In the subanalysis, this protective effect of the probiotics against diarrhea was observed only in trials that used whole milk (RR=0.154, 95% CI 0.079-0.301) and trials that used multistrain inocula (RR=0.415, 95% CI 0.227-0.759). Probiotics did not improve the fecal characteristics (standardized mean difference, SMD=-0.4904, 95% CI -1.011-0.035) and were unable to change the LAB:coliforms ratio (SMD=0.016, 95% CI -0.701-0.733). Probiotics showed a beneficial impact on the LAB:coliforms ratio in the subanalysis that included trials that used whole milk (SMD=0.780, 95% CI 0.141-1.418) and monostrain inocula (SMD=0.990, 95% CI 0.340-1.641). The probability of significant effects (probiotic positive effect) in a new study was >0.70 for diarrhea and fecal consistency. Whole milk feeding improved the action of the probiotic effect on the incidence of diarrhea and LAB:coliforms ratio. The probability to find significant effects in the diarrhea frequency and LAB:coliforms ratio was higher (P>0.85) if the new studies were conducted using whole milk to feed calves. This paper defines the guidelines to standardize the experimental designs of future trials. LAB can be used as growth promoters in calves instead of antibiotics to counteract the negative effects of their widespread use.

Evaluation of anti-Moraxella bovis pili immunoglobulin-A in tears following intranasal vaccination of cattle.

Infectious bovine keratoconjunctivitis (IBK) is a highly contagious ocular disease of cattle caused by Moraxella bovis (Mb). Parenterally administered immunogens used to prevent the disease do not offer complete protection possibly because they stimulate a poor ocular mucosal secretory response, in which locally secreted immunoglobulin-A (sIgA) is one of the main components. The principal aim of this study was to evaluate by an indirect enzyme linked immunosorbent assay (ELISA), the local ocular mucosal sIgA response against Mb purified pili, produced after intranasal inoculation of experimental vaccines. Pili were adjuvanted by several different adjuvants (QuilA, Marcol Arlacel, Marcol Span, microencapsulated pili with PLGA polymers). Results were compared to sIgA response produced by adjuvant placebo inoculations and by IBK natural infection. Significantly higher anti-pili IgA response (p<0.05) was detected in calves vaccinated intranasally with pili QuilA and pili Marcol Span compared to control calves, although this specific immune response did not seem to be related to protection against Mb infection or typical IBK lesion development.

Dynamics of Moraxella bovis infection and humoral immune response to bovine herpes virus type 1 during a natural outbreak of infectious bovine keratoconjunctivitis in beef calves.

Infectious bovine keratoconjunctivitis (IBK) is an acute disease caused by Moraxella bovis (Mb). Several factors may predispose animals to an IBK outbreak; one commonly observed is infection with bovine herpes virus type 1 (BHV-1). The aim of this study was to investigate the dynamics of BHV-1 virus infection and its relation with clinical cases of IBK in weaned calves from a beef herd with a high prevalence of lesions caused by Mb. Sampling was carried out in six stages and included conjunctival swabs for isolating Mb as well as blood samples for identifying antibodies specific for BHV-1. A score for IBK lesions after observing each eye was determined. The findings of this study showed a high prevalence of BHV-1 virus infection (100% of animals were infected at the end of the trial); 67% of animals were culture-positive for Mb, but low rates of clinical IBK (19% of calves affected) were detected at the end of the trial. These results suggest that infection with BHV-1 did not predispose these animals to IBK, and that Mb infection produced clinical and subclinical disease in the absence of BHV-1 co-infection.