Porcine intestinal epithelial cell lines as a new in vitro model for studying adherence and pathogenesis of enterotoxigenic Escherichia coli

Enterotoxigenic Escherichia coli (ETEC) infections result in large economic losses in the swine industry worldwide. The organism causes diarrhea by adhering to and colonizing enterocytes in the small intestines. While much progress has been made in understanding the pathogenesis of ETEC, no homologous intestinal epithelial cultures suitable for studying porcine ETEC pathogenesis have been described prior to this report. In the current study, we investigated the adherence of various porcine ETEC strains to two porcine (IPEC-1 and IPEC-J2) and one human (INT-407) small intestinal epithelial cell lines. Each cell line was assessed for its ability to support the adherence of E. coli expressing fimbrial adhesins K88ab, K88ac, K88ad, K99, F41, 987P, and F18. Wild-type ETEC expressing K88ab, K88ac, and K88ad efficiently bound to both IPEC-1 and IPEC-J2 cells. An ETEC strain expressing both K99 and F41 bound heavily to both porcine cell lines but an E. coli strain expressing only K99 bound very poorly to these cells. E. coli expressing F18 adhesin strongly bound to IPEC-1 cells but did not adhere to IPEC-J2 cells. The E. coli strains G58-1 and 711 which express no fimbrial adhesins and those that express 987P fimbriae failed to bind to either porcine cell line. Only strains B41 and K12:K99 bound in abundance to INT-407 cells. The binding of porcine ETEC to IPEC-J2, IPEC-1 and INT-407 with varying affinities, together with lack of binding of 987P ETEC and non-fimbriated E. coli strains, suggests strain-specific E. coli binding to these cell lines. These findings suggest the potential usefulness of porcine intestinal cell lines for studying ETEC pathogenesis.     

A proposal of clinical breakpoints for amoxicillin applicable to porcine respiratory tract pathogens

In the present position paper, an attempt was made to establish clinical breakpoints of amoxicillin to classify porcine respiratory tract pathogens as susceptible, intermediate or resistant based on their minimum inhibitory concentrations of amoxicillin. For this, a thorough review of the published literature with regard to swine-specific pharmacological data (including dosages of amoxicillin applied and routes of administration used), clinical efficacy, and in vitro susceptibility of the target pathogens was performed. Based on the comparative analysis of the results, the working group "Antibiotic Resistance" of the German Veterinary Medical Society (DVG) proposed to classify porcine respiratory tract pathogens that show MIC values of amoxicillin of < or =0.5microg/ml as "susceptible", those with MICs of 1microg/ml as "intermediate", and those with MICs of > or =2microg/ml as "resistant".     

A new PCR followed by MboI digestion for the detection of all variants of the Clostridium perfringens cpb2 gene

Clostridium perfringens which is a causative agent of several diseases in animals and humans is capable of producing a variety of toxins. Isolates are typed into five types on the basis of the presence of one or more of the four major toxins genes, i.e. cpa, cpb, etx, and iap. A decade ago another toxin termed beta2 (beta2) and its gene (cpb2) were identified. Two alleles of cpb2 are known and a possible link between differences in gene expression and allelic variation has been reported. A correlation between the level of expression and the origin of the isolates has also been suggested. The demonstration and typing of the cpb2 gene in the genome of isolates can be seen as a vital part of research on the role of the beta2 toxin in the pathogenesis of disease. This study describes a PCR with a single primer set which in contrast to published primer sets recognizes both alleles. Subsequent restriction enzyme analysis of the PCR product enables typing of the alleles. Applying this protocol on a total of 102 isolates, a sub-variant was found which occurred only in C. perfringens isolates from pigs and appeared to be the predominant variant found in C. perfringens isolates from this species.     

The effects of Strongylus vulgaris parasitism on eosinophil distribution and accumulation in equine large intestinal mucosa

REASONS FOR PERFORMING STUDY: Eosinophilic granulocytes have been associated with parasite or immune-mediated diseases, but their functions in other disease processes remain unclear. Cause and timing of eosinophil migration into the equine gastrointestinal mucosa are also unknown. OBJECTIVE: To determine the effects of intestinal parasitism on eosinophils in equine large intestinal mucosa. METHODS: Large intestinal mucosal samples were collected from horses and ponies (n = 16) from the general veterinary hospital population, ponies (n = 3) raised in a parasite-free environment, ponies experimentally infected with 500 infective Strongylus vulgaris larvae and treated with a proprietary anthelmintic drug (n = 14), and a similar group of ponies (n = 7) that received no anthelmintic treatment. Total eosinophil counts and eosinophil distribution in the mucosa were determined by histological examination. A mixed model analysis was performed and appropriate Bonferroni adjusted P values used for each family of comparisons. P<0.05 was considered significant. RESULTS: There was no difference in large intestinal mucosal eosinophil counts and eosinophil distribution between ponies infected with S. vulgaris and those raised in a parasite-free environment. Experimental infection with S. vulgaris, with or without subsequent anthelmintic treatment, did not change eosinophil counts, and counts were similar to those for horses from the general population. CONCLUSIONS: Migration of eosinophils to the equine large intestinal mucosa appears to be independent of exposure to parasites. Large intestinal mucosal eosinophils may have more functions in addition to their role in defence against parasites.     

The risk and control of Salmonella outbreaks in calf-raising operations: a mathematical modeling approach

Salmonellosis in calves has economic and welfare implications, and serves as a potential source of human infections. Our objectives were to assess the risk of Salmonella spread following its introduction into a herd of pre-weaned calves and to evaluate the efficacy of control strategies to prevent and control outbreaks. To meet these objectives, we developed a model of Salmonella transmission within a pre-weaned group of calves based on a well documented outbreak of salmonellosis in a calf-raising operation and other literature. Intervention scenarios were evaluated in both deterministic and stochastic versions of the model. While the basic reproduction number (R0) was estimated to be 2.4, simulation analysis showed that more than 60% of the invasions failed after the introduction of a single index case. With repeated introduction of index cases, the probability of Salmonella spread was close to 1, and the tested control strategies were insufficient to prevent transmission within the group. The most effective strategies to control ongoing outbreaks were to completely close the rearing operation to incoming calves, to increase the proportion of admitted calves that were immunized (>75%), and to assign personnel and equipment to groups of calves.     

Mineral content of hay harvested in Bavarian and Swiss horse farms. Predictive value of cutting time,number of cut,botanical composition,origin and fertilization

Hay samples from 29 horse farms in Southern Upper Bavaria and 31 horse farms in Switzerland were taken and analysed for minerals such as calcium, magnesium, phosphorus and potassium. The content of herbs and legumes in the hay was determined and the Weende analysis was performed. Comparison between Bavaria and Switzerland resulted in comparable calcium contents of approximately 4 g Ca/kg dry matter (DM) in grass hay. Hay with more than 10% of herbs and legumes found only in Switzerland showed higher Ca contents of >7 g Ca/kg DM in hay. The mineral contents of phosphorus and potassium were lower in hay from Switzerland (P: 1.8 ± 0.7 g/kg DM, K: 15.4 ± 5.1 g/kg DM) than from Bavaria (P: 3.8 ± 0.6 g/kg DM, K: 20.0 ± 6.0 DM), whereas the magnesium content of the hay showed no difference between the regions (∼1.5 g Mg/kg DM). Very late first grass hay cuts showed low magnesium and calcium (<4 g/kg DM) contents. Further minerals in the hay for horses differed from those in dairy cattle in the same region. Fertilization showed only small effects on the mineral contents of the hay produced for horses. Therefore, in calculation of ration and production of supplements for horses, these differences should be taken into account.     

Effects of B-trichothecenes on luminal glucose transport across the isolated jejunal epithelium of broiler chickens

Trichothecenes are closely-related sesquiterpenoids (ring structure) with a 12, 13 epoxy ring and a variable number of hydroxyl, acetyl or other substituents. In chickens, D-glucose and amino acid absorption occurs via carrier-mediated transport. Recently, it has been observed that deoxynivalenol (DON) alters the gut function and impairs glucose and amino acid transport in chickens. The purpose of this work was to determine the effects of different B-trichothecenes [DON, Nivalenol (NIV), 15-Ac-DON and Fusarenon X (FUS X)] on intestinal carrier-mediated sodium co-transport of D-glucose in the small intestine of broiler chickens. Intestinal transport was determined by changes in the short-circuit current (Isc), proportional to ion transmembrane flux, in the middle segment of the jejunum of broilers with the Ussing chamber technique. D-glucose produced an increase of the Isc, and this effect was reverted by different B-trichothecene mycotoxins, indicating that the glucose induced Isc was altered by B-trichothecenes. The addition of glucose after pre-incubation of the tissues with B-trichothecenes had no effect (p > 0.05) on the Isc, suggesting that B-trichothecenes afflicted the Na(+)-D-glucose co-transport. However, FUX had no obvious effect on the measured parameters. It could be concluded from the present study that the glucose co-transporter activity appears to be more sensitive to DON, NIV and 15-Ac-DON suppression than by FUS X in the jejunum of broilers.     

Experimental infection of domestic pigeons with pigeon circovirus

Pigeon circovirus (PiCV) infection and young pigeon disease syndrome (YPDS), associated with high morbidity and mortality, have been recognized in young racing pigeons from large portions of Central Europe. There exist a number of data indicating that YPDS is a consequence of PiCV infection and subsequent immunosuppression. In order to prove PiCV to be one of the crucial factors of YPDS, an experimental infection with PiCV was performed under controlled conditions. Twenty-four domestic pigeons (Columba livia forma domestica) were divided into two groups with 12 pigeons each; an infection group and a control group. All birds were between their fourth to eighth week of life. Pigeons in the infection group were infected both intramuscularly and orally with PiCV purified from naturally infected birds, while pigeons in the control group received a placebo. To test a possible influence of the PiCV infection on the immune system, the animals in both groups were vaccinated simultaneously, on the same day, against PMV-1 (Lasovac plus, IDT, Dessau-Tornau, Germany). Weekly virologic testing showed a viraemic period, and excretion of the infection virus, in pigeons in the infection group. Replication of PiCV could be proved on the basis of histologic findings of multiglobular inclusion bodies, mainly observed in macrophages of the bursa of Fabricius. A PiCV, genetically distinct from the experimental virus, was detected in the control group by polymerase chain reaction (PCR) testing, but any histologic findings comparable to the infection group were absent. None of the pigeons revealed clinical signs of illness, or hints that immunosuppression had occurred, regardless of their group. The absence of stressful conditions, considered as a trigger for the development of YPDS, may be responsible for the failure of disease reproduction in our infection model.     

Abrasions and lameness in piglets born in different farrowing systems with different types of floor

Background

The quality of the floor is essential to the welfare of piglets as abrasions often are recorded in newborn piglets, and such lesions may lead to lameness. Apart from animal suffering, lameness contributes to losses in form of dead piglets, decreased growth, and increased use of antibiotics and manual labour.

Methods

In a herd with three different farrowing systems, 37 litters (390 piglets) were studied until the age of 3 weeks with respect to presence of skin wounds and abrasions. Lameness was registered until the age of 7 weeks. Eight lame piglets were sacrificed before medical treatment and subjected to necropsy including histopathological and microbiological examinations. Isolates of streptococci, staphylococci and E. coli were tested with respect to antimicrobial resistance. Mastitis was observed in ten sows.

Results

The most severe abrasions at carpus and soles were seen in the system with a new solid concrete floor with a slatted floor over the dunging area. The lowest magnitude was observed in the deep litter system with peat. Sole bruising was more common in the systems with concrete floor compared to the deep litter system with peat, and the differce in prevalence was significant at all examination days. The lesions decreased with time and about 75% of the treatments for lameness were performed during the first three weeks of life. The overall prevalence of lameness was highest in the system with new solid concrete floor with a slatted floor over the dunging area (9.4%) followed by the old solid concrete floor (7.5%). A lower (p < 0.05) prevalence was seen in the deep litters system with peat (3.3%). No significant relationship between mastitis and abrasions or lameness in the offspring was observed.

Conclusion

There were large differences in the prevalence of abrasions and lameness between the floor types. The deep litter system with peat provided a soft and good floor for piglets. The overall prevalence of lameness was only diagnosed in every fourth litter in that system compared to in every second litter in the systems with concrete floor. In contrast, the incidence of mastitis in the sows during the first week after farrowing was higher than in the systems with concrete floor.