Heat stress and cow factors affect bacteria shedding pattern from naturally infected mammary gland quarters in dairy cattle.

Mastitis-causing pathogens are shed from infected mammary gland quarters and thus contribute to an increased risk of new intramammary infections. The objective of the current study was to investigate the shedding characteristics of various mastitis-causing pathogens and associated animal-specific (somatic cell score and parity) and environmental (heat stress) factors. In a longitudinal study, infected udder quarters were sampled consecutively on 5 dairy farms in Germany. To capture climatic factors, temperature-humidity index (THI) was calculated. In the laboratory analysis, the pathogens and their counts in the milk samples were determined. A generalized linear mixed model with gamma link was used to evaluate the factors influencing pathogen-shedding characteristics. The variables somatic cell count, pathogen, parity, and THI had significant influence on pathogen shedding. Staphylococci were shed in lower values than streptococci. The pathogen shedding from mammary gland quarters with intramammary infections was higher in the first and second lactation than in higher lactations. Exceeding the THI threshold 60 resulted in higher pathogen counts on the same day. This was only caused by the pathogens yeasts and Streptococcus uberis. Possible mechanisms causing differences in pathogen shedding are changes in the counts due to influenced milk quantities, better growth conditions at higher temperatures, or altered immunological reactions. The mechanisms often remain speculative and require further investigation. The study underlines the contribution of cows with high somatic cell counts regarding the transmission of mastitis pathogens within a herd. Furthermore, it becomes clear that heat stress in Germany influences udder health and that prevention measures are useful.

Cats shedding pathogenic Leptospira spp.-An underestimated zoonotic risk?

Shedding of DNA of pathogenic Leptospira spp. has been documented in naturally infected cats in several countries, but urinary shedding of infectious Leptospira spp. has only recently been proven. The climate in Southern Chile is temperate rainy with high annual precipitations which represents ideal preconditions for survival of Leptospira spp., especially during spring and summer. The aims of this study were to investigate shedding of pathogenic Leptospira spp. in outdoor cats in Southern Chile, to perform molecular characterization of isolates growing in culture, and to assess potential risk factors associated with shedding. Urine samples of 231 outdoor cats from rural and urban areas in southern Chile were collected. Urine samples were investigated for pathogenic Leptospira spp. by 4 techniques: qPCR targeting the lipL32 gene, immunomagnetic separation (IMS)-coupled qPCR (IMS-qPCR), direct culture and IMS-coupled culture. Positive urine cultures were additionally confirmed by PCR. Multilocus sequence typing (MLST) was used to molecularly characterize isolates obtained from positive cultures. Overall, 36 urine samples (15.6%, 95% confidence interval (CI) 11.4-20.9) showed positive results. Eighteen (7.8%, 95% CI 4.9-12.1), 30 (13%, 95% CI 9.2-18), 3 (1.3%, 0.3-3.9) and 4 cats (1.7%; 95% CI 0.5-4.5) were positive in qPCR, IMS-qPCR, conventional culture, and IMS-coupled culture, respectively. MLST results of 7 culture-positive cats revealed sequences that could be assigned to sequence type 17 (6 cats) and sequence type 27 (1 cat) corresponding to L. interrogans (Pathogenic Leptospira Subgroup 1). Shedding of pathogenic Leptospira spp. by cats might be an underestimated source of infection for other species including humans. The present study is the first one reporting growth of leptospires from feline urine in culture in naturally infected cats in South-America and characterisation of culture-derived isolates. So far, very few cases of successful attempts to culture leptospires from naturally infected cats are described worldwide.

A Gut Commensal-Produced Metabolite Mediates Colonization Resistance to Salmonella Infection.

The intestinal microbiota provides colonization resistance against pathogens, limiting pathogen expansion and transmission. These microbiota-mediated mechanisms were previously identified by observing loss of colonization resistance after antibiotic treatment or dietary changes, which severely disrupt microbiota communities. We identify a microbiota-mediated mechanism of colonization resistance against Salmonella enterica serovar Typhimurium (S. Typhimurium) by comparing high-complexity commensal communities with different levels of colonization resistance. Using inbred mouse strains with different infection dynamics and S. Typhimurium intestinal burdens, we demonstrate that Bacteroides species mediate colonization resistance against S. Typhimurium by producing the short-chain fatty acid propionate. Propionate directly inhibits pathogen growth in vitro by disrupting intracellular pH homeostasis, and chemically increasing intestinal propionate levels protects mice from S. Typhimurium. In addition, administering susceptible mice Bacteroides, but not a propionate-production mutant, confers resistance to S. Typhimurium. This work provides mechanistic understanding into the role of individualized microbial communities in host-to-host variability of pathogen transmission.

Quantifying fecal shedding of Mycobacterium avium ssp. paratuberculosis from calves after experimental infection and exposure.

Johne's disease, a chronic enteritis caused by Mycobacterium avium ssp. paratuberculosis (MAP), causes large economic losses to the dairy industry worldwide. Fecal shedding of MAP contaminates the environment, feed, and water and contributes to new infections on farm, yet there is limited knowledge regarding mechanisms of shedding, extent of intermittent shedding, and numbers of MAP bacteria shed. The objectives were to (1) compare (in an experimental setting) the frequency at which intermittent shedding occurred and the quantity of MAP shed among pen mates that were inoculated or contact-exposed (CE); and (2) determine whether an association existed between inoculation dose and quantity of MAP shed. In the first experiment, 32 newborn Holstein-Friesian bull calves were allocated to pens in groups of 4, whereby 2 calves were inoculated with a moderate dose (MD; 5 × 108 cfu) of MAP and 2 calves acted as CE. Calves were group-housed for 3 mo, fecal samples were collected and cultured, and culture-positive samples were quantified. In the second experiment, 6 calves were inoculated with either a low (LD) or high (HD) dose of MAP (1 × 108 or 1 × 1010 cfu, respectively), and fecal samples were collected for 3 mo and cultured for detection of MAP. The amount of MAP was quantified using direct extraction (DE) of DNA from fecal samples and F57-specific quantitative PCR. In experiment 1, the average amount of MAP in all culture-positive samples did not differ between MD and CE calves. In experiment 2, when comparing inoculation doses, LD calves had the lowest proportion of MAP-positive culture samples and HD had the highest, but no difference was detected in the average quantity of MAP shed. This study provided new information in regards to Johne's disease research and control regarding shedding from various inoculation doses and from CE animals; these data should inform future trials and control programs.

Changes in bacterial community composition of Escherichia coli O157:H7 super-shedder cattle occur in the lower intestine.

Escherichia coli O157:H7 is a foodborne pathogen that colonizes ruminants. Cattle are considered the primary reservoir of E. coli O157:H7 with super-shedders, defined as individuals excreting > 104 E. coli O157:H7 CFU g-1 feces. The mechanisms leading to the super-shedding condition are largely unknown. Here, we used 16S rRNA gene pyrosequencing to examine the composition of the fecal bacterial community in order to investigate changes in the bacterial microbiota at several locations along the digestive tract (from the duodenum to the rectal-anal junction) in 5 steers previously identified as super-shedders and 5 non-shedders. The overall bacterial community structure did not differ by E. coli O157:H7 shedding status; but several differences in the relative abundance of taxa and OTUs were noted between the two groups. The genus Prevotella was most enriched in the non-shedders while the genus Ruminococcus and the Bacteroidetes phylum were notably enriched in the super-shedders. There was greater bacterial diversity and richness in samples collected from the lower- as compared to the upper gastrointestinal tract (GI). The spiral colon was the only GI location that differed in terms of bacterial diversity between super-shedders and non-shedders. These findings reinforced linkages between E. coli O157:H7 colonization in cattle and the nature of the microbial community inhabiting the digestive tract of super-shedders.

Influence of On-farm pig Salmonella status on Salmonella Shedding at Slaughter.

The risk of Salmonella shedding among pigs at slaughter with regard to their previous on-farm Salmonella status was assessed in a group of pigs from a farm from NE of Spain. A total of 202 pigs that had been serologically monitored monthly during the fattening period and from which mesenteric lymph nodes (MLN) and faecal (SFEC) samples were collected at slaughter for Salmonella isolation were included. A repeated-measures anova was used to assess the relationship between mean OD% values during the fattening period and sampling time and bacteriology on MLN and SFEC. Pigs were also grouped into four groups, that is pigs seronegative during the fattening period and Salmonella negative in MLN (group A; n = 69); pigs seronegative during the fattening period but Salmonella positive in MLN (B; n = 36); pigs seropositive at least once and Salmonella positive in MLN (C; n = 50); and pigs seropositive at least once but Salmonella negative in (D; n = 47). Pigs shedding at slaughter seroconverted much earlier and showed much higher mean OD% values than non-shedders pigs. The proportion of Salmonella shedders in groups A and D was high and similar (26.1% and 29.8%, respectively), but significantly lower than that for groups B and C. The odds of shedding Salmonella for groups B and C were 4.8 (95% CI = 1.5-15.5) and 20.9 (3.7-118) times higher, respectively, when compared to A. It was concluded that a large proportion of Salmonella seronegative pigs may shed Salmonella at slaughter, which would be likely associated to previous exposure with contaminated environments (i.e. transport and lairage). For pigs already infected at farm, the likelihood of shedding Salmonella was much higher and may depend on whether the bacterium has colonized the MLN or not. The odds of shedding Salmonella spp. were always much higher for pigs in which Salmonella was isolated from MLN.

Infant Mouse Model for the Study of Shedding and Transmission during Streptococcus pneumoniae Monoinfection.

One of the least understood aspects of the bacterium Streptococcus pneumoniae (pneumococcus) is its transmission from host to host, the critical first step in both the carrier state and the disease state. To date, transmission models have depended on influenza A virus coinfection, which greatly enhances pneumococcal shedding to levels that allow acquisition by a new host. Here, we describe an infant mouse model that can be utilized to study pneumococcal colonization, shedding, and transmission during bacterial monoinfection. Using this model, we demonstrated that the level of bacterial shedding is highest in pups infected intranasally at age 4 days and peaks over the first 4 days postchallenge. Shedding results differed among isolates of five different pneumococcal types. Colonization density was found to be a major factor in the level of pneumococcal shedding and required expression of capsule. Transmission within a litter occurred when there was a high ratio of colonized "index" pups to uncolonized "contact" pups. Transmission was observed for each of the well-colonizing pneumococcal isolates, with the rate of transmission proportional to the level of shedding. This model can be used to examine bacterial and host factors that contribute to pneumococcal transmission without the effects of viral coinfection.

Broilers with low serum Mannose-binding Lectin show increased fecal shedding of Salmonella enterica serovar Montevideo.

Mannose-binding lectin (MBL) is a key molecule in innate immunity. MBL binds to carbohydrates on the surface of pathogens, initiating the complement system via the lectin-dependent pathway or facilitates opsonophagocytosis. In vivo studies using inbred chicken lines differing in MBL serum concentration indicate that chicken MBL affects Salmonella resistance; further studies are imperative in conventional broiler chickens. In this study 104 conventional day-old chickens (offspring from a cross between Cobb 500 male and female parent breeders) were orally infected with Salmonella enterica subsp. enterica serovar Montevideo. The chickens were divided into two groups based on polymorphisms in their MBL promoter region, designated L/L for low serum concentrations of MBL and L/H for medium serum concentrations of MBL. A semi-quantitative real-time PCR method for detection of Salmonella in cloacal swabs was used, the log10 CFU quantification was based on a standard curve from artificially spiked cloacal swab samples pre-incubated for 8 h with known concentrations of Salmonella ranging from 10(1) to 10(6) CFU/swabs, with an obtained amplification efficiency of 102% and a linear relationship between the log10 CFU and the threshold cycle Ct values of (R(2) = 0.99). The L/L chickens had significantly higher Log10 CFU/swab at week 5 post infection (pi) than the L/H chickens. A repetition of the study with 86 L/L and 18 L/H chickens, also gave significantly higher log10 CFU ± SEM in cloacal swabs, using the semi-quantitative real-time PCR method from L/L chickens than from the L/H chickens at week 5 pi. These results indicate that genetically determined basic levels of MBL may influence S. Montevideo susceptibility.

Feeding wet distillers grains plus solubles with and without a direct-fed microbial to determine performance, carcass characteristics, and fecal shedding of O157:H7 in feedlot heifers.

The inclusion of wet distillers grains plus solubles (WDGS) in feedlot diets has become a common practice in many regions of the United States due to the expanded production of byproducts and fluctuating corn prices related to ethanol production and other factors. In addition, societal concerns over the continued use of antimicrobials in agriculture production combined with an enhanced interest in disease and pathogen prevention in the food supply have led to an increased interest in use of direct-fed microbials (DFM) in growing and finishing cattle. Direct-fed microbials have been shown to improve ADG and feed efficiency, alter ruminal fermentation, and decrease fecal shedding of potential harmful pathogens in feedlot cattle in some experiments. The objective of this experiment was to evaluate the effects of WDGS inclusion with or without a DFM containing (1 × 10 cfu ∙ heifer ∙ d) combined with (1 × 10 cfu ∙ heifer ∙ d) on the performance, carcass characteristics, and O157:H7 shedding in feedlot heifers. In early August, 288 crossbred heifers (initial BW = 295 ± 28 kg) were assigned to 1 of 4 treatments (12 pens per treatment; 6 heifers per pen) in a randomized complete block design with a 2 × 2 factorial arrangement of treatments. Body weights and fecal grab samples were obtained at approximately 28-d intervals throughout the experiment. Across the feeding period, heifers fed 30% WDGS tended ( = 0.09) to have greater ADG and had greater carcass-adjusted ADG ( = 0.05) compared with heifers fed dry-rolled corn (DRC). Dry matter intake was not affected ( = 0.65) by diet, although carcass-adjusted G:F tended ( = 0.10) to be improved for heifers fed WDGS. Heifers fed 30% WDGS tended ( ≤ 0.10) to have greater fat thickness at the 12th rib, lower marbling scores, and higher yield grades. The inclusion of . combined with . in the diet had no effect ( > 0.10) on performance or carcass merit in the present experiment. The incidence of O157:H7 throughout the experiment was low, with only 18 positive samples across all sampling periods. Neither WDGS inclusion nor the inclusion of . combined with . in the diet had any effect ( > 0.10) on O157:H7 shedding in this experiment. Feeding 30% WDGS to feedlot heifers improved animal performance compared to the DRC-based control diet.

Patterns in Leptospira Shedding in Norway Rats (Rattus norvegicus) from Brazilian Slum Communities at High Risk of Disease Transmission.

BACKGROUND: We address some critical but unknown parameters of individuals and populations of Norway rats (Rattus norvegicus) that influence leptospiral infection, maintenance and spirochetal loads shed in urine, which contaminates the environment ultimately leading to human infection. METHODOLOGY/PRINCIPAL FINDINGS: Our study, conducted in Salvador, Brazil, established the average load of leptospires in positive kidneys to be 5.9 x 10(6) per mL (range 3.1-8.2 x10(6)) genome equivalents (GEq), similar to the 6.1 x 10(6) per ml (range 2.2-9.4 x10(6)) average obtained from paired urines, with a significant positive correlation (R2=0.78) between the two. Based on bivariate and multivariate modeling, we found with both kidney and urine samples that leptospiral loads increased with the age of rats (based on the index of body length to mass), MAT titer and the presence of wounding/scars, and varied with site of capture. Some associations were modified by sex but trends were apparent. Combining with data on the demographic properties and prevalence of leptospiral carriage in rat populations in Salvador, we estimated that daily leptospiral loads shed in the urine of a population of 82 individuals exceeded 9.1 x 10(10) leptospires. CONCLUSIONS/SIGNIFICANCE: These factors directly influence the risk of leptospiral acquisition among humans and provide essential epidemiological information linking properties of rat populations with risk of human infection.

Contagious epididymitis due to Brucella ovis: relationship between sexual function, serology and bacterial shedding in semen.

BACKGROUND: Contagious Epididymitis (CE) due to Brucella ovis (B. ovis) is a contagious disease that impairs rams' fertility due to epididymis, testicle and accessory sexual gland alterations. An increased incidence of CE has been observed in South Eastern France ("PACA" region) since the Rev.1 vaccination against B. melitensis has been stopped in 2008. The objective of this study was to evaluate the relationship between the infection by B. ovis and the sexual function of rams. Two-hundred eighteen sexually-mature rams, from 11 seropositive flocks, were submitted to a clinical examination of the genital tract, a semen collection by electro-ejaculation for spermogram and culture, and a serological examination for anti-B. ovis antibodies by complement fixation test (CFT) and indirect ELISA (I-ELISA). The relationships between clinical, seminal, bacteriological and serological parameters were studied using the Fisher exact test and a logistic regression model (binomial logit). RESULTS: B. ovis shedding in semen was significantly associated with seropositivity (CFT and I-ELISA; p < 0.001 and 0.01 respectively), genital tract alterations (p < 0.05) and poor semen quality (p < 0.001). Seropositive rams presented significantly more genital tract alterations (p < 0.001) and a poor seminal score (p < 0.001) than seronegative rams. CONCLUSIONS: Since semen culture is not routinely feasible in field conditions, a control plan of CE should be based, where Rev.1 vaccination is not possible, on both systematic clinical and serological examination of rams, followed by the culling of seropositive and/or genital tract alterations carrier rams.

The use of direct-fed microbials to reduce shedding of Escherichia coli O157 in beef cattle: a systematic review and meta-analysis.

Human illness due to infections with Escherichia coli O157 is a serious health concern. Infection occurs through direct contact with infected animals or their faeces, through contaminated food or water and/or through person-to-person transmission. A reduction in faecal E. coli O157 shedding in cattle might reduce the burden of human infections. We used systematic review and meta-analysis to assess the efficacy of direct-fed microbials (DFM), compared with placebo or no treatment, fed during the pre-harvest stage of production in reducing faecal E. coli O157 shedding in beef cattle during field trials. Four electronic databases, Nebraska Beef Reports and review article reference lists were searched. A total of 16 publications assessing faecal shedding at the end of the trial and/or throughout the trial period were included. The majority of publicly disseminated trials evaluated the prevalence of E. coli O157 faecal shedding; only two evaluated the concentration of organisms in faeces. The prevalence of faecal E. coli O157 shedding in cattle is significantly reduced by DFM treatments (summary effect size for all DFM - OR = 0.46; CI = 0.36-0.60). The DFM combination Lactobacillus acidophilus (NP51) and Propionibacterium freudenreichii (NP24) was more efficacious in reducing the prevalence of faecal E. coli O157 shedding at the time of harvest and throughout the trial period compared with the group of other DFM, although this difference was not statistically significant. Furthermore, we found that the combination [NP51 and NP24] treatment was more efficacious in reducing the prevalence of faecal E. coli O157 shedding at the time of harvest and throughout the trial period when fed at the dose of 10(9) CFU/animal/day than any lesser amount, although this difference was not statistically significant. Feeding beef cattle DFM during the pre-harvest stage of production reduces the prevalence of E. coli O157 faecal shedding and might effectively reduce human infections.

Fate of non O157 Shigatoxigenic Escherichia coli in ovine manure composting / Eliminação de Escherichia coli Shigatoxigênica não O157 em compostagem de esterco ovino

Livestock manure may contain pathogenic microorganisms which pose a risk to the health of animal or humans if the manure is not adequately treated or disposed of. To determine the fate of Shiga toxigenic Escherichia coli (STEC) non O157 in composted manure from naturally colonized sheep, fresh manure was obtained from animals carrying bacterial cells with stx1/ stx2 genes. Two composting systems were used, aerated and non-aerated, and the experiments were done in Dracena city, São Paulo State. Every week, for seven weeks, one manure sample from six different points in both systems was collected and cultured to determine the presence of E. coli, the presence of the virulence genes in the cells, and also the susceptibility to 10 antimicrobial drugs. The temperature was verified at each sampling. STEC non-O157 survived for 49 days in both composting systems. E. coli non-STEC showing a high degree of antibiotic resistance was recovered all long the composting period. No relationship was established between the presence of virulence genes and antibiotic resistance. The presence of virulence genes and multiple antibiotic resistances in E. coli implicates a potential risk for these genes spread in the human food chain, which is a reason for concern...

Esterco de animais de criação pode conter microrganismos patogênicos, o que representa um risco para a saúde animal e a humana se o esterco não for adequadamente tratado ou descartado. Determinou-se o tempo necessário para a eliminação de Escherichia coli Shiga toxigenica (STEC) não O157 em esterco ovino composto, obtido de fezes frescas de ovelhas naturalmente colonizadas com cepas STEC não O157 que apresentavam os genes stx1/ stx2. Foram utilizados dois sistemas de compostagem, aerado e não aerado, em experimentos realizados na cidade de Dracena, estado de São Paulo. Todas as semanas, durante sete semanas, uma amostra de compostagem proveniente de seis pontos diferentes na leira, nos dois sistemas, foi coletada e semeada para a determinação da presença de E. coli, da presença de genes de virulência nas células, bem como da sensibilidade dessas células a 10 drogas antimicrobianas. Em cada amostragem, a temperatura da leira foi analisada. Células de STEC não O157 sobreviveram por 49 dias nos dois sistemas de compostagem. E. coli não STEC com um alto grau de resistência a antibióticos foi recuperada ao longo de todo o período de compostagem. Não foi possível estabelecer relação entre a presença de genes de virulência e a resistência a antibióticos. A presença de genes de virulência e a resistência a múltiplos antibióticos em E. coli representam um risco potencial para o espalhamento desses genes na cadeia alimentar humana, o que é motivo de grande preocupação...

TLR2 signaling decreases transmission of Streptococcus pneumoniae by limiting bacterial shedding in an infant mouse Influenza A co-infection model.

While the importance of transmission of pathogens is widely accepted, there is currently little mechanistic understanding of this process. Nasal carriage of Streptococcus pneumoniae (the pneumococcus) is common in humans, especially in early childhood, and is a prerequisite for the development of disease and transmission among hosts. In this study, we adapted an infant mouse model to elucidate host determinants of transmission of S. pneumoniae from inoculated index mice to uninfected contact mice. In the context of co-infection with influenza A virus, the pneumococcus was transmitted among wildtype littermates, with approximately half of the contact mice acquiring colonization. Mice deficient for TLR2 were colonized to a similar density but transmitted S. pneumoniae more efficiently (100% transmission) than wildtype animals and showed decreased expression of interferon α and higher viral titers. The greater viral burden in tlr2-/- mice correlated with heightened inflammation, and was responsible for an increase in bacterial shedding from the mouse nose. The role of TLR2 signaling was confirmed by intranasal treatment of wildtype mice with the agonist Pam3Cys, which decreased inflammation and reduced bacterial shedding and transmission. Taken together, these results suggest that the innate immune response to influenza virus promotes bacterial shedding, allowing the bacteria to transit from host to host. These findings provide insight into the role of host factors in the increased pneumococcal carriage rates seen during flu season and contribute to our overall understanding of pathogen transmission.

Evaluation of animal genetic and physiological factors that affect the prevalence of Escherichia coli O157 in cattle.

Controlling the prevalence of Escherichia coli O157 in cattle at the pre-harvest level is critical to reduce outbreaks of this pathogen in humans. Multilayers of factors including the environmental and bacterial factors modulate the colonization and persistence of E. coli O157 in cattle that serve as a reservoir of this pathogen. Here, we report animal factors contributing to the prevalence of E. coli O157 in cattle. We observe the lowest number of E. coli O157 in Brahman breed when compared with other crosses in an Angus-Brahman multibreed herd, and bulls excrete more E. coli O157 than steers in the pens where cattle were housed together. The presence of super-shedders, cattle excreting >10(5) CFU/rectal anal swab, increases the concentration of E. coli O157 in the pens; thereby super-shedders enhance transmission of this pathogen among cattle. Molecular subtyping analysis reveal only one subtype of E. coli O157 in the multibreed herd, indicating the variance in the levels of E. coli O157 in cattle is influenced by animal factors. Furthermore, strain tracking after relocation of the cattle to a commercial feedlot reveals farm-to-farm transmission of E. coli O157, likely via super-shedders. Our results reveal high risk factors in the prevalence of E. coli O157 in cattle whereby animal genetic and physiological factors influence whether this pathogen can persist in cattle at high concentration, providing insights to intervene this pathogen at the pre-harvest level.

Coxiella burnetii shedding during the peripartum period and subsequent fertility in dairy cattle.

The objective of this study was to assess the effects of Coxiella burnetii shedding or seropositivity on post-partum recovery and subsequent fertility in high-producing dairy cows. Given the difficulty in diagnosing C. burnetii infection at the farm level, an exhaustive series of tests in 43 pregnant animals that delivered at least one live calf were conducted, including blood serology and PCR of milk or colostrum, cotyledons (only at parturition), faeces, vaginal fluid against C. burnetii on gestation Day 171-177, at parturition and on Days 1-7, 8-14, 15-21, 22-28, 29-35 and 90-97 post-partum. During scheduled herd visits, ultrasonography (US) of the genital tract and examination of vaginal fluid were performed on Days 15-21 (V1), 22-28 (V2), 29-35 (V3) and 51-57 (V4) post-partum by the same veterinarian. Logistic regression analysis revealed that the likelihood of suffering endometritis (the presence of echogenic intrauterine fluid (IUF), cervical diameter of ≥4 cm or endometrial thickness ≥0.75 cm) was lower in C. burnetii-seropositive animals (OR = 0.10), compared with C. burnetii-seronegative animals. According to Kaplan-Meier survival analysis, C. burnetii-seronegative and non-shedding cows showed a delayed return to luteal activity and conception was delayed in non-shedding animals, compared with the remaining animals. Overall, the results of our study provide useful insight into the effects of C. burnetii infection on post-partum recovery and subsequent fertility. In particular, animals not infected with Coxiella seem to be susceptible to infection and not protected against the bacterium in dairy herds. The elevated costs of determining an infection at the farm level, make monitoring of cows virtually impossible from a clinical point of view.

Effect of organic acids on Salmonella colonization and shedding in weaned piglets in a seeder model.

Piglets (n = 128) weaned at 21 days of age were used in a 35-day seeder model to evaluate the effects of dietary additives differing in active ingredients, chemical, and physical formulation, and dose on Salmonella colonization and shedding and intestinal microbial populations. Treatments were a negative control (basal diet), the positive control (challenged, basal diet), and six treatments similar to the positive control but supplemented with the following active ingredients (dose excluding essential oils or natural extracts): triglycerides with butyric acid (1.30 g kg(-1)); formic and citric acids and essential oils (2.44 g kg(-1)); coated formic, coated sorbic, and benzoic acids (2.70 g kg(-1)); salts of formic, sorbic, acetic, and propionic acids, their free acids, and natural extracts (2.92 g kg(-1)); triglycerides with caproic and caprylic acids and coated oregano oil (1.80 g kg(-1)); and caproic, caprylic, lauric, and lactic acids (1.91 g kg(-1)). On day 6, half the piglets (seeder pigs) in each group were orally challenged with a Salmonella Typhimurium nalidixic acid-resistant strain (4 × 10(9) and 1.2 × 10(9) log CFU per pig in replicate experiments 1 and 2, respectively). Two days later, they were transferred to pens with an equal number of contact pigs. Salmonella shedding was determined 2 days after challenge exposure and then on a weekly basis. On day 34 or 35, piglets were euthanized to sample tonsils, ileocecal lymph nodes, and ileal and cecal digesta contents. The two additives, both containing short-chain fatty acids and one of them also containing benzoic acid and the other one also containing essential oils, and supplemented at more than 2.70 g kg(-1), showed evidence of reducing Salmonella fecal shedding and numbers of coliforms and Salmonella in cecal digesta. However, colonization of tonsils and ileocecal lymph nodes by Salmonella was not affected. Supplementing butyric acid and medium-chain fatty acids at the applied dose failed to inhibit Salmonella contamination in the current experimental setup.

The strain-specific dynamics of Escherichia coli O157:H7 faecal shedding in cattle post inoculation.

This study reports analysis of faecal shedding dynamics in cattle for three Escherichia coli O157:H7 (ECO157) strains (S1, S2 and S3) of different genotype and ecological history, using experimental inoculation data. The three strains were compared for their shedding frequency and level of ECO157 in faeces. A multistate Markov chain model was used to compare shedding patterns of S1 and S2. Strains S1 and S2 were detected seven to eight times more often and at 10(4) larger levels than strain S3. Strains S1 and S2 had similar frequencies and levels of shedding. However, the total time spent in the shedding state during colonization was on average four times longer for S1 (15 days) compared to S2 (4 days). These results indicate that an ECO157 strain effect on the frequency, level, pattern and the duration of faecal shedding may need to be considered in control of ECO157 in the cattle reservoir.

Cost-effectiveness of diagnostic strategies to identify Mycobacterium avium subspecies paratuberculosis super-shedder cows in a large dairy herd using antibody enzyme-linked immunosorbent assays, quantitative real-time polymerase chain reaction, and bacterial culture.

Diagnostic strategies to detect Mycobacterium avium subsp. paratuberculosis (MAP) super-shedder cows in dairy herds have been minimally studied. The objective of the current study was to compare the cost-effectiveness of strategies for identification of MAP super-shedders on a California dairy herd of 3,577 cows housed in free-stall pens. Eleven strategies that included serum or milk enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qPCR) or culture of environmental samples, pooled or individual cow fecal samples, or combinations thereof were compared. Nineteen super-shedders (0.5%) were identified by qPCR and confirmed by culture as cows shedding ≥ 10,000 colony forming units (CFU)/g feces (median of 30,000 CFU/g feces). A stratified random sample of the study herd based on qPCR results of fecal pools was the most sensitive (74%) strategy and had the highest cost ($5,398/super-shedder). The reference strategy with the lowest cost ($1,230/super-shedder) and sensitivity (47%) included qPCR testing of fecal samples from ELISA-positive lactating (milk) and nonlactating (serum) cows housed in pens with the highest MAP bioburden. The most cost-effective alternative to the reference was to perform qPCR testing of fecal samples from ELISA-positive cows (milk and serum for milking and dry cows, respectively) for a sensitivity of 68% and cost of $2,226/super-shedder. In conclusion, diagnostic strategies varied in their cost-effectiveness depending on the tests, specimen type, and labor costs. Initial qPCR testing of environmental samples from free-stall pens to target cows in pens with the highest MAP bioburden for further testing can improve the cost-effectiveness of strategies for super-shedder identification.

Salmonella enterica shedding in hospitalized horses and associations with diarrhea occurrence among their stablemates and gastrointestinal-related illness or death following discharge.

OBJECTIVE: To evaluate the potential association between Salmonella enterica shedding in hospitalized horses and the risk of diarrhea among stablemates, and to characterize gastrointestinal-related illness and death following discharge among horses that shed S. enterica while hospitalized. DESIGN: Retrospective cohort study [corrected]. ANIMALS: 221 horses (59 that shed S. enterica during hospitalization and 162 that tested negative for S. enterica shedding ≥ 3 times during hospitalization). PROCEDURES: Information from medical records (signalment, results of microbial culture of fecal samples, clinical status at the time of culture, and treatment history) was combined with data collected through interviews with horse owners regarding formerly hospitalized horses and their stablemates. Data were analyzed to investigate risk factors for death and diarrhea. RESULTS: Occurrence of diarrhea among stablemates of formerly hospitalized horses was not associated with S. enterica shedding in hospitalized horses but was associated with oral treatment with antimicrobials during hospitalization. Salmonella enterica shedding during hospitalization was not associated with risk of death or gastrointestinal-related illness in study horses ≤ 6 months after discharge, but shedding status and history of gastrointestinal illness were associated with increased risk of death during the preinterview period. CONCLUSIONS AND CLINICAL RELEVANCE: Stablemates of horses that shed S. enterica during hospitalization did not appear to have an increased risk for diarrhea, but comingling with horses that receive orally administered antimicrobials may affect this risk. Salmonella enterica shedding during hospitalization may be a marker of increased long-term risk of death after discharge. Risks are likely influenced by the S enterica strain involved and biosecurity procedures used.