Virulence profiles of enterotoxigenic, shiga toxin and enteroaggregative Escherichia coli in South African pigs

Enterotoxigenic Escherichia coli (ETEC) and shiga toxin E. coli (STEC) are important causes of colibacillosis in piglets. Recently, enteroaggregative E. coli heat-stable enterotoxin 1 (EAST-1) has been implicated in pig diarrhoea. This study investigated the prevalence of enterotoxin [heat-labile toxins (LT), heat-stable toxin a (STa), heat-stable toxin b (STb)], shiga toxins (Stx1, Stx2, Stx2e), enteroaggregative heat-stable E. coli (EAST-1), associated fimbriae (F4, F5, F6, F41, F18ab, F18ac) and non-fimbrial adhesins [adhesin involved in diffuse adherence 1 (AIDA-1), attaching and effacing factor, porcine attaching- and effacing-associated factor] in South African pigs. A total of 263 E. coli strains were isolated from Landrace (n?=?24), Large White (n?=?126), Duroc (n?=?28) and indigenous (n?=?85) breeds of piglets aged between 9 and 136 days. PCR was used in the analysis. Virulent genes were detected in 40.3 % of the isolates, of which 18.6, 0.4 and 17.5 % were classified as ETEC, STEC and enteroaggregative E. coli (EAEC), respectively. Individual genes were found in the following proportions: STb (19.01 %), LT (0.4 %), STa (3.4 %), St2xe (1.1 %) and EAST-1 (20.2 %) toxins. None of the tested fimbriae were detected in ETEC and STEC isolates. About one third of the ETEC and STEC isolates was tested negative for both fimbrial and non-fimbrial adhesins. Twenty-five pathotypes from ETEC-, EAEC- and STEC-positive strains were identified. Pathotypes EAST-1 (30.2 %), STb (13.2 %) and STb/AIDA-1 (10.4 %) were most prevalent. The study provided insight on possible causes of colibacillosis in South African pigs.     

Both flagella and F4 fimbriae from F4ac+ enterotoxigenic Escherichia coli contribute to attachment to IPEC-J2 cells in vitro

The role of flagella in the pathogenesis of F4ac⁺ Enterotoxigenic Escherichia coli (ETEC) mediated neonatal and post-weaning diarrhea (PWD) is not currently understood. We targeted the reference {"type":"entrez-nucleotide","attrs":{"text":"C83902","term_id":"2706834","term_text":"C83902"}}C83902 ETEC strain (O8:H19:F4ac⁺ LT⁺ STa⁺ STb⁺), to construct isogenic mutants in the fliC (encoding the major flagellin protein), motA (encoding the flagella motor), and faeG (encoding the major subunit of F4 fimbriae) genes. Both the ΔfliC and ΔfaeG mutants had a reduced ability to adhere to porcine intestinal epithelial IPEC-J2 cells. F4 fimbriae expression was significantly down-regulated after deleting fliC, which revealed that co-regulation exists between flagella and F4 fimbriae. However, there was no difference in adhesion between the ΔmotA mutant and its parent strain. These data demonstrate that both flagella and F4 fimbriae are required for efficient F4ac⁺ ETEC adhesion in vitro.     

Protective effect of oral administration of transgenic tobacco seeds against verocytotoxic Escherichia coli strain in piglets

The use of transgenic plants as delivery system for antigenic proteins is attractive for its simplicity and increases likelihood for local immune response at sites of infection. The aim of this study was to evaluate the protective effect of oral administration of tobacco seeds, expressing the FedA, the major protein of the F18 adhesive fimbriae, and B subunit of verocytotoxin, against verocytotoxin-producing E. coli (VTEC) strain in piglets. Forty-three early weaned piglets, were randomly divided into 4 experimental groups: 3 test groups and a control. Treatment groups orally received a bolus, with different dose of tobacco seeds on 0, 1, 2, 14 days post primary administration. After challenge, with 1*10¹⁰ CFU of O138 Escherichia coli strain, piglets showed clinical scores significantly higher in the control group compared to orally immunized groups (P?<?0.05) and the latter showed a faster recovery than in CG. In conclusion, oral administration of recombinant tobacco seeds expressing antigenic proteins against VTEC strains can induce a protective effect against challenger strain in piglets.     

Prevalence,prediction and risk factors of enteropathogens in normal and non-normal faeces of young Dutch dairy calves

Between January and April 2007, 424 calves under 22 days of age from 108 Dutch dairy herds were sampled to estimate the prevalence of non-normal faeces (‘custard-like’—yellowish-coloured with custard consistency or diarrhoea: watery-like faeces) and the shedding of enteropathogens Escherichia coli K99 (E. coli), Coronavirus, Cryptosporidium parvum (C. parvum), Rotavirus and Clostridium perfringens (Cl. perfringens). In addition, information was collected on animal characteristics and herd-management practices. The probability of detecting each one of five enteropathogens given a calf with ‘custard-like’ faeces or diarrhoea was estimated using Bayes’ rule and was based on the predicted probabilities from a multinominal model including each of five enteropathogens as independent variables. In addition, putative risk factors for the presence of each of five enteropathogens were analysed using logistic regression models with random herd effects.Fifty-seven percent of calves had faeces of normal colour (brownish) and consistency (firm), 23.8% (95%CI: 19.8–28.2%) had ‘custard-like’ faeces and 19.1% (95%CI: 15.5–23.2%) had diarrhoea. E. coli was the least detected enteropathogen (2.6% (95%CI: 1.3–4.6%) of calves, 9% (95%CI: 5–16%) of herds) and Cl. perfringens was most detected (54.0% (95%CI: 49.1–58.8%) of calves, 85% (95%CI: 77–91%) of herds). E. coli and Coronavirus were detected incidentally in only one or two calves per herd, whereas C. parvum and Cl. perfringens were frequently detected in up to four calves per herd. For calves with ‘custard-like’ faeces, the probability of detecting Rotavirus from a calf in its first week of age was 0.31 whereas for a calf in its second week, there was a 0.66 probability of detecting C. parvum. The probabilities of detecting E. coli, Rotavirus and C. parvum in calves with diarrhoea in their first week of age were 0.10, 0.20 and 0.43, respectively. In calves with diarrhoea between 1 and 2 weeks of age, the probability of detecting enteropathogens was 0.43 for C. parvum. None of the tested enteropathogens were related to ‘custard-like’ faeces or diarrhoea in the third week of age.Putative risk factors for E. coli, Coronavirus and C. parvum included the presence of peer-calves shedding Coronavirus, C. parvum or Rotavirus, respectively. Additionally, managerial risk factors such as non-optimal hygienic housing (for Coronavirus) and the routine use of antibiotics for diarrhoeic calves (for C. parvum) were found. No animal or managerial factors were associated with shedding of Cl. perfringens.     

Early supplementation with Lactobacillus plantarum in liquid diet modulates intestinal innate immunity through toll-like receptor 4-mediated mitogen-activated protein kinase signaling pathways in young piglets challenged with Escherichia coli K88

This study was conducted to investigate the effects of early supplementation during 4 to 18 d of age with Lactobacillus plantarum (LP) in liquid diets on intestinal innate immune response in young piglets infected with enterotoxigenic Escherichia coli (ETEC) K88. Seventy-two barrow piglets at 4 d old were assigned to basal or LP-supplemented liquid diet (5 × 10¹⁰ CFU·kg⁻¹). On day 15, piglets from each group were orally challenged with either ETEC K88 (1 × 10⁸ CFU·kg⁻¹) or the same amount of phosphate-buffered saline. The intestinal mucosa, mesenteric lymph node (MLN), and spleen samples were collected on day 18. Here, we found that LP pretreatment significantly decreased the mRNA relative expression of inflammatory cytokines (interleukin [IL]-1β, IL-8, and tumor necrosis factor-α), porcine β-defensin 2 (pBD-2), and mucins (MUC1 and MUC4) in the jejunal mucosa in piglets challenged with ETEC K88 (P < 0.05). Moreover, LP significantly decreased the ileal mucosa mRNA relative expression of IL-8 and MUC4 in young piglets challenged with ETEC K88 (P < 0.05). Furthermore, the piglets of the LP + ETEC K88 group had lower protein levels of IL-8, secretory immunoglobulin A, pBD-2, and MUC4 in the jejunal mucosa than those challenged with ETEC K88 (P < 0.05). Besides, LP supplementation reduced the percentage of gamma/delta T cells receptor (γδTCR) and CD172a⁺ (SWC3⁺) cells in MLN and the percentage of γδTCR cells in the spleen of young piglets after the ETEC K88 challenge. Supplementation with LP in liquid diets prevented the upregulated protein abundance of toll-like receptor (TLR) 4, phosphorylation-p38, and phosphorylation-extracellular signal-regulated protein kinases in the jejunal mucosa induced by ETEC K88 (P < 0.05). In conclusion, LP supplementation in liquid diet possesses anti-inflammatory activity and modulates the intestinal innate immunity during the early life of young piglets challenged with ETEC K88, which might be attributed to the suppression of TLR4-mediated mitogen-activated protein kinase signaling pathways. Early supplementation with LP in liquid diets regulates the innate immune response, representing a promising immunoregulation strategy for maintaining intestinal health in weaned piglets.     

The survey of porcine teschoviruses, porcine circovirus and porcine transmissible gastroenteritis virus infecting piglets in clinical specimens in China

A multiplex PCR assay was developed and evaluated for its ability to simultaneously detect three viral infections of swine. Specific primers were carefully selected from articles published for each of the following three viruses: porcine circovirus type II (PCV2), porcine teschovirus (PTV) and porcine transmissible gastroenteritis virus (TGEV). Each target produced a specific amplicon with a size of 353 bp (PCV2), 168 bp (PTV) and 499 bp (TGEV). The sensitivity of the multiplex PCR using purified plasmid constructs containing the specific viral target fragments was 6.60?×?10², 8.43?×?10² and 7.30?×?10² copies for PCV2, PTV and TGEV, respectively. Among 127 samples which were collected from Heilongjiang, Jilin, Henan and Guangxi provinces, the single infection of PCV2, PTV and TGEV was 99.21, 46.88 and 65.35 %, respectively, and co-infection of the three viruses was 26.77 %. In conclusion, the multiplex PCR has the potential to be useful for routine molecular diagnosis and epidemiology.     

Prevalence of fimbial colonization factors F18ab and F18ac in Escherichia coli isolates from weaned piglets with edema and/or diarrhea in China

F18ab and F18ac are important fimbrial colonization factors of verotoxigenic Escherichia coli (VTEC) and/or enterotoxigenic E. coli (ETEC) in weaned piglets with edema disease and/or diarrhea. To further investigate their prevalence and correlation to pathogenic E. coli, a duplex PCR, using three primers derived from the nucleotide sequence of the F18 major fimbrial subunit gene (fedA), and a direct agglutination test, using a monoclonal antibody specific for the antigenic factor 'a' of F18, were performed. Among 60VTEC, 24VTEC/ETEC and 24 ETEC isolates tested from weaned piglets with edema disease and/or diarrhea in different pig farms in the Jiangsu Province of China, 52 isolates (48.15%) were positive in the direct agglutination test and 63 isolates (58.33%) were positive in the duplex PCR. Among 63 PCR-positive isolates, 53 isolates (49.07%) were F18ab-positive and 10 isolates (9.26%) were F18ac-positive. In addition, the F18ab gene was more frequently detected in VTEC (61.67%) or VTEC/ETEC (62.50%) than in ETEC (4.17% only), while the F18ac gene was more frequently detected in VTEC/ETEC (33.33%) than in ETEC (8.33%) or VTEC (0%). Furthermore, F18ab was more frequently associated with Shiga toxin 2e (Stx2e), whereas F18ac was more frequently associated with enterotoxin ST I. These results suggest that the duplex PCR performed in this experiment is a more reliable method for identification of F18+E. coli, and that F18 is a more important virulence factor of VTEC and VTEC/ETEC.     

Neonatal Diarrhea of Pigs in Quebec: Infectious Causes of Significant Outbreaks

To evaluate the relative importance of the various enteropathogens causing neonatal diarrhea in Quebec farrowing operations, observations were made on 749 diarrheic pigs from 325 outbreaks of diarrhea. They were one to 15 days of age, and were obtained alive for necropsy generally within 48 hours of the onset of diarrhea. Some pigs were from severe, explosive outbreaks of diarrhea with high morbidity and mortality rates, while others were from herds with chronic neonatal diarrhea with lower morbidity and mortality rates. A combination of bacteriological, virological and histological methods were used to study the pigs.

Viruses were incriminated in 60%, bacteria in 23% and coccidia in 15.3% of the 325 diarrhea outbreaks. Transmissible gastroenteritis virus was by far the most common enteropathogen with a prevalence of 52%; rotavirus was implicated in 9.2% of the outbreaks while adenovirus was incriminated in 0.30% of the outbreaks. Enterotoxigenic Escherichia coli were involved in 22.4% of the cases while Clostridium perfringens type C was an occasional finding. Coccidia involved in our herds were identified as Isospora suis. The disease was attributed to infection with a single etiologic agent in 590 diarrheic pigs (78%) while combinations of agents were present in only 90 (12%).

The age-specific occurrence of the various enteropathogens was evaluated. Transmissible gastroenteritis virus was the most common enteropathogen in all age groups. Colibacillosis was common in pigs which became diarrheic under five days of age; in this age group, the enterotoxigenic E. coli were frequently found alone, but were usually combined with other agents in older pigs. The prevalence of coccidia was high in pigs which became diarrheic between five and 15 days of age. Rotavirus infection was common in diarrheic pigs older than ten days of age. Although individual baby pigs were commonly infected with a single enteropathogen, it was very common to see more than one agent involved in an outbreak of diarrhea, particularly when pigs of different ages were affected.

Observations on the occurrence of the enteropathogens according to the seasons were also made. Occurrence of transmissible gastroenteritis was throughout the year with the highest prevalence during the fall, winter and spring months. Colibacillosis and coccidiosis were more common in the summer, fall and early winter months with the lowest prevalence in the spring months.

     

Inverse relationship between heat stable enterotoxin-b induced fluid accumulation and adherence of F4ac-positive enterotoxigenic Escherichia coli in ligated jejunal loops of F4ab/ac fimbria receptor-positive swine

Heat-labile enterotoxin (LT) produced by enterotoxigenic Escherichia coli (ETEC) increases bacterial adherence to porcine enterocytes in vitro and enhances small intestinal colonization in swine. Heat-stable enterotoxin-b (STb) is not known to affect colonization; however, through an induction of net fluid accumulation it might reduce bacterial adherence. The relationship between fluid accumulation and bacterial adherence in jejunal loops inoculated with ETEC strains that produce LT, STb, both, or neither toxin was studied. Ligated jejunal loops were constructed in weaned Yorkshire pigs in two independent experiments (Exp. 1, n = 5, 8-week-old; Exp. 2, n = 6, 6–8-week-old). Each pig was inoculated with six F4ac⁺ E. coli strains: (1) LT⁺, STb⁺ parent (WAM2317); (2) STb^? (ΔestB) mutant (MUN297); (3) MUN297 complemented with STb (MUN298); (4) LT^? STb^? (ΔeltAB ΔestB) mutant (MUN300); (5) MUN300 complemented with LT (MUN301); and (6) 1836-2 (non-enterotoxigenic, wild-type). Pigs were confirmed to be K88 (F4)ab/ac receptor-positive in Exp. 2 by testing for intestinal mucin-type glycoproteins and inferred to be receptor-positive in both Exp. 1 and 2 based on histopathologic evidence of bacterial adherence. Strains that produced STb induced marked fluid accumulation with the response (ml/cm) to WAM2317 and MUN298 significantly greater than that to the other strains (P < 0.0001). Conversely, bacterial adherence scores based on immunohistochemistry and CFU/g of washed mucosa were both lowest in the strains that expressed STb and highest in those that did not. For the two experiments combined, the Pearson correlation coefficient (R) between fluid volume (ml/cm) and log CFU per gram was ?0.57021 (P < 0.0001); R² = 0.3521 (n = 197). These results support the hypothesis that enterotoxin-induced fluid accumulation flushes progeny organisms into the lumen of the bowel, thereby increasing the likelihood of fecal shedding and transmission of the pathogen to new hosts.     

An investigation of the effects of intratesticular alfaxalone and lidocaine during castration in piglets

Objective

To determine whether intratesticular injection of an alfaxalone–lidocaine combination can induce anesthesia and provide a rapid recovery in piglets undergoing surgical castration.

An investigation into the association between cpb2-encoding Clostridium perfringens type A and diarrhea in neonatal piglets

To investigate the possible role of cpb2-positive type A Clostridium perfringens in neonatal diarrheal illness in pigs, the jejunum and colon of matched normal and diarrheic piglets from 10 farms with a history of neonatal diarrhea were examined grossly and by histopathology, and tested for C. perfringens, for C. perfringens beta2 (CPB2) toxin, as well as for Clostridium difficile toxins, Salmonella, enterotoxigenic Escherichia coli, rotavirus, transmissible gastroenteritis (TGE) virus, and coccidia. Clostridium perfringens isolates were tested using a multiplex real-time polymerase chain reaction (PCR) to determine the presence of cpa, consensus and atypical cpb2, and other virulence-associated genes. The numbers of C. perfringens in the intestinal contents were lower in diarrheic piglets (log₁₀ 5.4 CFU/g) compared with normal piglets (log₁₀ 6.5 CFU/g) (P < 0.05). The consensus cpb2 was present in 93% of isolates in each group, but atypical cpb2 was less common (56% healthy, 32% diarrheic piglets isolates, respectively, P < 0.05). The presence of CPB2 toxin in the intestinal contents of normal and diarrheic piglets did not differ significantly. Clostridium difficile toxins and rotavirus were each detected in 7 of the 21 (33%) diarrheic piglets. Rotavirus, C. difficile toxins, Salmonella, or enterotoxigenic E. coli were concurrently recovered in different combinations in 4 diarrheic piglets. The cause of diarrhea in 8 of the 21 (38%) piglets on 6 farms remained unknown. The etiological diagnosis of diarrhea could not be determined in any of the piglets on 2 of the farms. This study demonstrated that the number of cpb2-positive type A C. perfringens in the intestinal contents was not a useful approach for making a diagnosis of type A C. perfringens enteritis in piglets. Further work is required to confirm whether cpb2-carrying type A C. perfringens have a pathogenic role in enteric infection in neonatal swine.     

High prevalence of F4<Superscript>+</Superscript> and F18<Superscript>+</Superscript><Emphasis Type="Italic">Escherichia coli</Emphasis> in Cuban piggeries as determined by serological survey

Little information is available on the prevalence of swine enteropathogens in Cuba where diarrheic diseases are responsible for 31% and 37% of the total mortality during the neonatal and postweaning periods. F4⁺ and F18⁺ enterotoxigenic Escherichia coli and F18⁺ verotoxigenic E. coli induce diarrhea and edematous disease in pigs, but their distribution has never been thoroughly studied in the Cuban swine population. Therefore, the present study estimated the prevalence of F4- and F18-specific antibodies in sera of 1,044 6-month-old gilts distributed in 34 piggeries spread over the Cuban territory. For the data analysis, which included the optical density of individual samples tested by ELISA, random-effects models and a mixture model in R (package “mixAK”; Komárek, Computational Statistics and Data Analysis 53:3932–3947, 2009) were fitted. Low, moderate, and high levels of F4-specific antibodies were found in 67.6%, 26.8%, and 5.6% of the gilts, while 66.4% and 33.6% of gilts showed low and high levels of F18-specific antibodies. Hereby, we show that F4⁺ and F18⁺ E. coli are highly prevalent as potential enteropathogens in Cuban piggeries.     

Quantitative real‐time PCR monitoring of Escherichia coli and Clostridium perfringens with oral administration of Lactobacillus plantarum strain Lq80 to weaning piglets

Levels of fecal or intestinal lactobacilli, Escherichia coli and Clostridium perfringens, and the prevalence of clostridial alpha toxin gene and heat‐stable toxin (ST) gene of enterotoxigenic E. coli (ETEC) were monitored in weaned piglets before (day 0) and during (days 7, 14, and 21) the administration of Lactobacillus plantarum strain Lq80. Lactobacilli were enumerated in a culture‐dependent method. The remainders were determined by quantitative real‐time PCR. In this quantitative real‐time PCR method, the detection limit was proved to be as low as 10³ cells/g feces or intestinal contents. Number of lactobacilli increased from day 0 to day 7 (P < 0.05), to day 14 (P < 0.05), and to day 21 (P = 0.07) in the Lq80‐administered group. L. plantarum contributed to as low as 10% of the lactobacillal population in the Lq80‐administered group. The number of E. coli and C. perfringens, and the prevalence of alpha toxin gene in feces or intestinal contents of the Lq80‐administered group decreased, at least in the first week of the postweaning period. Oral administration of L. plantarum strain Lq80 can stimulate the growth of indigenous lactobacilli and decrease ST‐producing ETEC and C. perfringens in the intestine of postweaning piglets.     

Genotyping and antimicrobial susceptibility of Clostridium perfringens isolated from dromedary camels,pastures and herders

The present study aimed to isolate and genotype C. perfringens from healthy and diarrheic dromedary camels, pastures and herders; and to evaluate and compare antimicrobial susceptibility of the isolates. A total of 262 (56.3%) C. perfringens isolates were recovered from 465 samples of healthy and diarrheic dromedary camels, pastures and herders. C. perfringens type A (75.2%), type B (4.2%), type C (13.7%) and type D (6.9%) were detected. C. perfringens type A with only cpa⁺ gene was found in 191 (72.9%) isolates and with cpa⁺ associated cpb2⁺ was found only in 6 (2.3%) isolates. None of the isolates were positive for cpe and iap genes.The highest antimicrobial resistance (82.8%) was observed to ceftiofur with MIC₅₀ and MIC₉₀ values of <64 and ≥256 μg/mL, respectively, followed by penicillin G (72.9%) and erythromycin (61.5%). The lowest resistance (1.9%) was observed for doxycycline with MIC₅₀ and MIC₉₀ values of <1 and 4 μg/mL, respectively, followed by florfenicol (5.3%) and clindamycin (12.2%). In conclusion, C. perfringens type A with cpa⁺ gene was the most prevalent toxin type isolated in this study. The majority of the isolates were resistant to at least one of the ten antimicrobials tested. Antimicrobial resistance patterns of C. perfringens isolates provide further evidence on the emergence of multiple-drug resistant C. perfringens. Therefore, the dissemination of surveillance programs to monitor and control C. perfringens in dromedary camels is required.     

Micro-encapsulated essential oils and organic acids combination improves intestinal barrier function,inflammatory responses and microbiota of weaned piglets challenged with enterotoxigenic Escherichia coli F4 (K88+)

This study evaluated the effects of micro-encapsulated (protected) organic acids (OA) and essential oils (EO) combination, P(OA + EO), and effects of a regular blend of free acids (FA) on the growth, immune responses, intestinal barrier and microbiota of weaned piglets challenged with enterotoxigenic Escherichia coli (ETEC) F4 (K88⁺). A total of 30 crossbred (Duroc × Landrace × Large White) weaned barrows (7.41 ± 0.06 kg, 28 d old) were assigned randomly to 5 treatments: 1) non-challenged positive control (PC), 2) ETEC F4 (K88⁺)-challenged negative control (NC), 3) NC + kitasamycin at 50 mg/kg + olaquindox at 100 mg/kg + free acidifier (FA) at 5 g/kg, 4) NC + kitasamycin at 50 mg/kg + olaquindox at 100 mg/kg + P(OA + EO) at 1 g/kg (P1), 5) NC + kitasamycin at 50 mg/kg + olaquindox at 100 mg/kg + P(OA + EO) at 2 g/kg (P2). Each dietary treatment had 6 replicates of one piglet each and the study lasted for 3 wk. On d 7, pigs in NC, FA, P1 and P2 were orally dosed with 10 mL of ETEC F4 (K88⁺) culture (1 × 10⁹ CFU/mL). From d 7 to 14 after the ETEC F4 (K88⁺) challenge, P1 increased gain-to-feed ratio (G:F) significantly (P < 0.05) compared with NC and FA groups. From d 14 to 21, P2 increased the average daily gain of pigs (P < 0.05) compared with NC and FA groups. Compared with NC, P2 reduced tumor necrosis factor-α (TNF-α), interleukin (IL)-6 and IL-10 concentrations (P < 0.05) in sera collected at 4 h later after ETEC F4 (K88⁺) challenge. On d 21, P1 increased occludin and zonula occludens-1 protein expression in ileum compared with NC (P < 0.05). After this 3-wk experiment, alpha diversity of gut microbiota was decreased by P2 compared with PC, and P1 increased the relative abundance of Lactobacillus in ileum, cecum and colon (P < 0.05). In conclusion, dietary P(OA + EO) additive at 2 g/kg combined with antibiotics could improve piglet performance and attenuate inflammation, and P(OA + EO) additive at 1 g/kg combined with antibiotics improved intestinal barrier and increased beneficial microbiota composition after an F4 (K88⁺) challenge.     

Studies on preweaning piglet diarrhoea

SUMMARY The aim of this study was to determine the cause and risk factors Involved in preweaning piglet diarrhoea. Faecal samples from 2380 diarrhoeic piglets, 5 to 30 days of age, were examined for enteropathogens. Isospora suls oocysts were detected in 53.8% of samples, Escherichia coll in 18.2% and rotavirus in 16.9%. I suis had the widest distribution, being present on 70.9% of 151 piggeries. The onset of diarrhoea occurred mainly between 7 and 14 days (77.5%), peaking at 10 days. Records of 4086 litters from two intensive piggeries were analysed and showed no seasonal variation or effect of sow parity on the incidence of piglet diarrhoea. I suis was the most common enteropathogen associated with diarrhoea in piglets from 5 days of age until weaning.     

Fibre supplementation to pre-weaning piglet diets did not improve the resilience towards a post-weaning enterotoxigenic E. coli challenge

Dietary fibre (DF) is implicated in gastrointestinal health of weaned piglets, either through its physiochemical properties, through modulation of gut microbiota and (or) improved gut integrity. We aimed to study the effect of DF enriched supplemental diets fed to suckling piglets (‘creep feed’) on health and performance after weaning when challenged with an enterotoxigenic E. coli (ETEC). Seventy-two piglets originating from 28 litters had been fed four creep diets, that is a low-fibre control (CON); a diet containing 2% long-chain arabinoxylans from wheat (lc-AXOS) or 5% purified cellulose (CELL) or a diet containing the high fermentable and the low-fermentable fibre source (i.e. 2% lc-AXOS and 5% CELL). Upon weaning, piglets were individually housed and all fed the same diet. On days 7, 8 and 9, animals received an oral dose of ETEC (5 ml containing 10⁷ to 10⁸ CFU/ml). Besides growth performance, faecal and skin scores were recorded daily. Gut permeability was assessed by urinary excretion of Co-EDTA prior and post-ETEC challenge. Repeated measures in time were statistically evaluated with generalized linear mixed models. We used a binominal distribution for evaluating the faecal and skin scores. Feed intake and body weight gain did not differ between treatments (p > .05). Piglets on CELL decreased gain:feed ratio in week 2 + 3 week compared to CON (p = .035). Prior to ETEC challenge, gut permeability tended to increase for lc-AXOS (p = .092). Moreover, lc-AXOS as main effect increased intestinal permeability before ETEC challenge (p = .013), whereas the low-fermentable fibre lead to elevated intestinal permeability after ETEC challenge (p = .014). The incidence of diarrhoea was higher for lc-AXOS + CELL compared with lc-AXOS (p = .036), while skin condition was unaffected. In conclusion, neither the high fermentable nor the low-fermentable fibre source improved post-weaning growth or gastrointestinal health of the piglets.     

Toxigenic Escherichia coli isolated from pigs in Argentina

The presence of porcine toxigenic E. coli (ETEC, VTEC) in 28 piggeries (5% of total) of the central and northeast region of Argentina was studied for a better understanding of the epidemiology of porcine strains. Samples were taken by rectal swabs from healthy piglets and from those with diarrhoea, in addition to their dams. Between 5-10 colonies were isolated from each one of 223 animals sampled from 1992 to 1997. By using specific primers each strain was screened by PCR for VT1, VT2all, VT2e, STIa, and LTI toxin genes. Only strains positive for any of the toxins mentioned above were screened for STb. Their O serogroups were determined by agglutination. All of the above enterotoxins and verocytotoxins were found in E. coli isolated from the animals. The STIa gene was detected in E. coli isolated from 27/127 piglets with diarrhoea, in comparison with LTI (4/127 pigs). No toxin gene was amplified from E. coli isolated from either healthy piglets or their dams. When strains isolated from 48 piglets without diarrhoea but showing delayed growth were analysed by PCR, their toxin profile was determined to be VT1 (1/48 piglets), VT2all (5/48), STIa (1/48), LTI (3/48) and VT2e (3/48). Serogroup O64 prevailed among ETEC; O138 prevailed for ETEC/VTEC strains. This is the first extensive study regarding porcine toxigenic E. coli in Argentina and constitutes an important database for the implementation of prevention measures.     

Pathology of calves with diarrhoea in southern Britain

Twenty-one moribund calves with diarrhoea were purchased from 11 farms, their faeces examined for enteropathogens and samples of intestinal tissue removed under anaesthesia. Lesions and presence of enteropathogens on the mucosal surface were scored by histological examination of immunostained paraffin sections. Two or more enteropathogens were detected in 19 calves. Cryptosporidium appeared to be the principal cause of diarrhoea in six calves, rotavirus in four, Salmonella typhimurium in two, bacteria adherent to the surface of the large intestine in two, coronavirus in one and K99+ Escherichia coli in one calf. Diarrhoea in four calves was the consequence of mixed infections in which no one enteropathogen appeared to predominate. In one calf no enteropathogen was detected. Diarrhoea was associated with infections and lesions throughout the small and large intestines. The enteropathogens most frequently associated with lesions in the small intestines were rotavirus, coronavirus and cryptosporidium; in the large intestines they were coronavirus and bacteria apparently adherent to the mucosal surface.