Diarrhea In Neonatal Piglets Research Articles

Characterization and Pathogenicity of the Porcine Deltacoronavirus Isolated in Southwest China.

Porcine deltacoronavirus (PDCoV) is a newly emerging enteric pathogen in swine that causes diarrhea in neonatal piglets and creates an additional economic burden on porcine industries in Asia and North America. In this study, a PDCoV isolate, CHN-SC2015, was isolated from Sichuan Province in southwest China. The isolate was characterized by a cytopathic effect, immunofluorescence, and electron microscopy. CHN-SC2015 titers in LLC-PK cells ranged from 104.31 to 108.22 TCID50/mL during the first 30 passages. During serial passage, 11 nucleotide mutations occurred in the S gene, resulting in nine amino acid changes. A whole genome sequencing analysis demonstrated that CHN-SC2015 shares 97.5%–99.1% identity with 59 reference strains in GenBank. Furthermore, CHN-SC2015 contained 6-nt deletion and 9-nt insertion in the ORF1ab gene, 3-nt deletion in the S gene and 11-nt deletion in its 3′UTR compared with other reference strains available in GenBank. A phylogenetic analysis showed that CHN-SC2015 is more closely related to other PDCoV strains in China than to the strains from Southeast Asia, USA, Japan, and South Korea, indicating the diversity of genetic relationships and regional and epidemic characteristics among these strains. A recombination analysis indicated that CHN-SC2015 experienced recombination events between SHJS/SL/2016 and TT-1115. In vivo infection demonstrated that CHN-SC2015 is highly pathogenic to sucking piglets, causing diarrhea, vomiting, dehydration, and death. Virus was shed daily in the feces of infected piglets and upon necropsy, was found distributed in the gastrointestinal tract and in multiple organs. CHN-SC2015 is the first systematically characterized strain from southwest China hitherto reported. Our results enrich the body of information on the epidemiology, pathogenicity and molecular evolution associated with PDCoV.

Antimicrobial resistance in production animals. Can homeopathy offer solutions? – Homeopathy as replacement to antibiotics in the case of neonatal diarrhoea in piglets

The use of antibiotics in the livestock sector is increasing to such an extent that it can lead to negative aspects for human health, animal health and the environment. Homeopathy might be an alternative to antibiotics and has therefore been tested in a randomized placebo-controlled trail to prevent Escherichia coli diarrhoea in neonatal piglets. On a commercial pig farm 52 sows of different parities, in their last month of gestation, were treated twice a week with either the homeopathic agent Coli 30K or placebo. The 525 piglets that were born from these sows were scored for occurrence and duration of diarrhoea. Piglets from the homeopathic treated group had significant (P = 0.0012 on pig-level and P < 0.05 on sow-level) less E. coli diarrhoea than piglets in the placebo group. The occurrence of diarrhoea seemed to be less severe in the homeopathic treated litters, where the disease did not spread further and duration tended to be shorter. Especially piglets from first parity sows gave a good response to treatment with Coli 30K. Homeopathic treatments in the livestock sector, in this case to prevent diarrhoea, could be a solution to lower the current use of veterinary antibiotics.

Functional Characterization and Proteomic Analysis of Porcine Deltacoronavirus Accessory Protein NS7.

Porcine deltacoronavirus (PDCoV) is an emerging swine enteric coronavirus that causes diarrhea in neonatal piglets. Like other coronaviruses, PDCoV encodes at least three accessory or species-specific proteins; however, the biological roles of these proteins in PDCoV replication remain undetermined. As a first step toward understanding the biology of the PDCoV accessory proteins, we established a stable porcine cell line constitutively expressing the PDCoV NS7 protein in order to investigate the functional characteristics of NS7 for viral replication. Confocal microscopy and subcellular fractionation revealed that the NS7 protein was extensively distributed in the mitochondria. Proteomic analysis was then conducted to assess the expression dynamics of the host proteins in the PDCoV NS7-expressing cells. Highresolution two-dimensional gel electrophoresis initially identified 48 protein spots which were differentially expressed in the presence of NS7. Seven of these spots, including two upregulated and five down-regulated protein spots, showed statistically significant alterations, and were selected for subsequent protein identification. The affected cellular proteins identified in this study were classified into functional groups involved in various cellular processes such as cytoskeleton networks and cell communication, metabolism, and protein biosynthesis. A substantial down-regulation of α-actinin-4 was confirmed in NS7-expressing and PDCoV-infected cells. These proteomic data will provide insights into the understanding of specific cellular responses to the accessory protein during PDCoV infection.

Stage of Gestation at Porcine Epidemic Diarrhea Virus Infection of Pregnant Swine Impacts Maternal Immunity and Lactogenic Immune Protection of Neonatal Suckling Piglets.

During pregnancy, the maternal immune response changes dramatically over the course of gestation. This has implications for generation of lactogenic immunity and subsequent protection in suckling neonates against enteric viral infections. For example, porcine epidemic diarrhea virus (PEDV) is an alphacoronavirus that causes acute diarrhea in neonatal piglets. Due to the high virulence of PEDV and the naïve, immature immune system of neonatal suckling piglets, passive lactogenic immunity to PEDV induced during pregnancy, via the gut-mammary gland (MG)-secretory IgA (sIgA) axis, is critical for piglet protection. However, the anti-PEDV immune response during pregnancy and stage of gestation required to optimally stimulate the gut-MG-sIgA axis is undefined. We hypothesize that there is a gestational window in which non-lethal PEDV infection of pregnant gilts influences maximum lymphocyte mucosal trafficking to the MG, resulting in optimal passive lactogenic protection in suckling piglets. To understand how the stages of gestation affect maternal immune responses to PEDV, three groups of gilts were orally infected with PEDV in the first, second or third trimester. Control (mock) gilts were inoculated with medium in the third trimester. To determine if lactogenic immunity correlated with protection, all piglets were PEDV-challenged at 3–5 days postpartum. PEDV infection of gilts at different stages of gestation significantly affected multiple maternal systemic immune parameters prepartum, including cytokines, B cells, PEDV antibodies (Abs), and PEDV antibody secreting cells (ASCs). Pregnant second trimester gilts had significantly higher levels of circulating PEDV IgA and IgG Abs and ASCs and PEDV virus neutralizing (VN) Abs post PEDV infection. Coinciding with the significantly higher PEDV Ab responses in second trimester gilts, the survival rate of their PEDV-challenged piglets was 100%, compared with 87.2, 55.9, and 5.7% for first, third, and mock litters, respectively. Additionally, piglet survival positively correlated with PEDV IgA Abs and ASCs and VN Abs in milk and PEDV IgA and IgG Abs in piglet serum. Our findings have implications for gestational timing of oral attenuated PEDV maternal vaccines, whereby PEDV intestinal infection in the second trimester optimally stimulated the gut-MG-sIgA axis resulting in 100% lactogenic immune protection in suckling piglets.

Isolation and characterization of a highly pathogenic strain of Porcine enteric alphacoronavirus causing watery diarrhoea and high mortality in newborn piglets.

Porcine enteric alphacoronavirus (PEAV) was first discovered in China in February 2017, and the origin and virulence of this novel porcine coronavirus were not fully characterized. Here, we isolated a strain of PEAV, named GDS04 that is identified by immunofluorescence and typical crown‐shaped particles observed with electron microscopy. Genomic analysis reveals that PEAV GDS04 shares a close relationship with SADS‐CoV and SeACoV. Furthermore, newborn piglets orally challenged with PEAV GDS04 developed typical clinical symptoms as watery diarrhoea in neonatal piglets. Viral RNA was detected in faeces and various tissues of the infected piglets. Moreover, macroscopic and microscopic lesions in whole intestinal tract were observed, and viral antigen could be detected in the small intestines by immunohistochemical staining and electron microscopy. Importantly, the mortality rate of inoculated‐newborn piglets was 100% and half of the cohabiting piglets died. Collectively, we demonstrate that PEAV is highly pathogenic in newborn piglets.

Diarrhoea in neonatal piglets: a case control study on microbiological findings

BackgroundMany factors can influence the occurrence of neonatal diarrhoea in piglets. Currently, well-known pathogens such as enterotoxigenic Escherichia coli and Clostridium perfringens type C appear to play a minor role in development of disease. Other infectious pathogens may be involved. In this study, we aimed to investigate the presence of selected infectious pathogens in neonatal piglets with clinical and pathological signs of enteric disease. The association between rotavirus A, Enterococcus hirae, Clostridium difficile and Clostridium perfringens type A/C and diarrhoea was investigated in a case control study on piglet level. The possible role of E. coli virulence factors was investigated in a multistep-procedure using herd-pools of E.coli isolates to screen for their presence.ResultsRotavirus A was detected more often in cases (25%) than in controls (6%) (P < 0.001). The detection rate of Enterococcus hirae, Clostridium difficile and C. perfringens type A positive for beta2 genes was the same in the two groups of piglets. C. perfringens type C was not detected in the study. Investigations on E. coli virulence factors showed a high prevalence of EAST1 toxin genes (55% of tested case piglets were positive) and AIDA-1 adhesin genes (63% of toxin positive case piglets were positive) in case piglets.ConclusionsDetection of rotavirus A was statistically significantly associated with neonatal piglet diarrhoea.An aetiologic role of E. coli carrying virulence factors EAST1 and AIDA-1 needs further investigation as the study points out these two factors as possible causative factors in neonatal diarrhoea.Detection of E.hirae, C.difficile and C. perfringens type A carrying beta 2 genes was not associated with neonatal piglet diarrhoea. However, the study suggested that massive overgrowth by E. hirae could be part of the pathogenesis in some cases of neonatal diarrhoea.

High prevalence of Clostridium difficile PCR ribotype 078 in pigs in Korea

Clostridium difficile can cause neonatal enteritis, but has been isolated from both diseased and healthy pigs. C. difficile shedding by pigs is a potential source of zoonotic transmission to humans. The objectives of this study were to characterize the prevalence and genotype of C. difficile with respect to age and health status in the pig industry for the first time in Korea. Fecal samples of 910 pigs were analyzed for the presence of C. difficile. In total, 176 (19.3%) C. difficile strains were isolated. The prevalence was significantly higher (p < 0.01) in diarrheic samples (30.4%) than in non-diarrheic samples (13.3%). The prevalence was highest in diarrheic suckling piglets (53.6%), followed by diarrheic sows (40.0%) and non-diarrheic suckling piglets (34.0%). The toxin genes tcdA, tcdB, cdtA, and cdtB were all detected in 153 isolates (A+B+CDT+, 86.9%). Furthermore, the majority of toxigenic strains (86.5%) were identified as PCR ribotype 078, which is an important pathogen in both pigs and humans. These results indicated that C. difficile is a leading causative pathogen of neonatal diarrhea in piglets in Korea.

Skin Vaccination against Rotavirus Using Microneedles: Proof of Concept in Gnotobiotic Piglets.

Live-attenuated oral rotavirus (RV) vaccines have lower efficacy in low income countries, and additionally are associated with a rare but severe adverse event, intussusception. We have been pursuing the development of an inactivated rotavirus vaccine (IRV) using the human rotavirus strain CDC-9 (G1P[8]) through parenteral immunization and previously demonstrated dose sparing and enhanced immunogenicity of intradermal (ID) unadjuvanted IRV using a coated microneedle patch in comparison with intramuscular (IM) administration in mice. The aim of this study was to evaluate the immune response and protection against RV infection and diarrhea conferred by the administration of the ID unadjuvanted IRV using the microneedle device MicronJet600® in neonatal gnotobiotic (Gn) piglets challenged with virulent Wa G1P[8] human RV. Three doses of 5 μg IRV when administered intradermally and 5 μg IRV formulated with aluminum hydroxide [Al(OH)3] when administered intramuscularly induced comparable rotavirus-specific antibody titers of IgA, IgG, IgG avidity index and neutralizing activity in sera of neonatal piglets. Both IRV vaccination regimens protected against RV antigen shedding in stools, and reduced the cumulative diarrhea scores in the piglets. This study demonstrated that the ID and IM administrations of IRV are immunogenic and protective against RV-induced diarrhea in neonatal piglets. Our findings highlight the potential value of an adjuvant sparing effect of the IRV ID delivery route.

Multidrug resistance in Clostridium perfringens isolated from diarrheal neonatal piglets in Thailand

Clostridium perfringens causes diarrhea in neonatal piglets, thereby affecting commercial swine farming. The objective of this study was to determine the prevalence and characterize antimicrobial resistance in C. perfringens isolated from diarrheal neonatal piglets in Thailand. A total of 260 rectal swab samples were collected from 13 farms and were subjected to C. perfringens isolation. A total of 148 samples were PCR-positive for C. perfringens toxin genes, from which 122 were recovered. All isolates were cpb2-encoding C. perfringens type A and enterotoxin gene negative. Most of the isolates were susceptible to ampicillin, bacitracin, chlorotetracycline, doxycycline, and oxytetracycline with MIC50 values ranging from 0.32 to 8 μg/ml. The high resistance rates were observed for ceftiofur, enrofloxacin, erythromycin, lincomycin, and tylosin. Among resistant isolates, 82% were resistant to more than one type of antibiotics. The distinct pattern of multiple drug resistance in C. perfringens was observed in different regions, potentially reflecting the farm specific usage of these agents.

A food-grade fimbrial adhesin FaeG expression system in Lactococcus lactis and Lactobacillus casei.

Enterotoxigenic Escherichia coli (ETEC) infection is the major cause of diarrhea in neonatal piglets. The fimbriae as colonizing factor in the pathogenesis of ETEC constitute a primary target for vaccination against ETEC. Lactic acid bacteria (LAB) are attractive tools to deliver antigens at the mucosal level. With the safety of genetically modified LAB in mind, a food-grade secretion vector (pALRc or pALRb) was constructed with DNA entirely from LAB, including the replicon, promoter, signal peptide, and selection marker alanine racemase gene (alr). To evaluate the feasibility of the system, the nuclease gene (nuc) from Staphylococcus aureus was used as a reporter to be expressed in both Lactococcus lactis and Lactobacillus casei. Subsequently, the extracellular secretion of the fimbrial adhesin FaeG of ETEC was confirmed by Western blot analysis. These results showed that this food-grade expression system has potential as the delivery vehicle for the safe use of genetically modified LAB for the development of vaccines against ETEC infection.

USO DE ÓLEOS ESSENCIAIS NO CONTROLE DOS SINAIS CLÍNICOS DAS DIARREIAS NEONATAIS EM LEITÕES NASCIDOS DE FÊMEAS COM DIFERENTES ORDENS DE PARTO

Este estudo avaliou a emulsão líquida de óleos essenciais de orégano (Origanum vulgare) e alecrim (Rosmarinus officinalis labiatae) no controle dos sinais clínicos da diarreia neonatal em leitões. Foram monitorados 1.334 leitões lactentes, com peso vivo médio inicial de 1,5kg, quanto aos sinais clínicos de diarreia neonatal. O delineamento experimental foi o inteiramente casualizado e com três tratamentos (T): (T1) controle (solução fisiológica), (T2) mescla dos óleos essenciais e (T3) enrofloxacina. Os animais que receberam o T1 apresentaram sinais clínicos característicos de diarreia diferindo (P&lt;0,01) dos demais tratamentos. O percentual de cura da diarreia dos leitões não diferiu (P&gt;0,01) entre os tratamentos, independente da ordem de partos (OP) das mães. Já leitões com sinais clínicos de diarreia, nascidos de fêmeas com OP inferior a três apresentaram maior percentual de cura (74,6%), se comparados aos leitões diarreicos de fêmeas com OP entre três e cinco (68,2%) e superior a cinco (56,4%). As análises microbiológicas das fezes e histológicas do epitélio ileal não apresentaram diferenças (P&gt;0,01) entre os tratamentos. A emulsão com óleos essenciais de orégano e alecrim (2mL kg-1), para leitões entre zero e o oito dias de vida, pode vir a substituir a enrofloxacina no controle dos sinais clínicos das diarreias neonatais.

Effect of a single dose of Saccharomyces cerevisiae var. boulardii on the occurrence of porcine neonatal diarrhoea

Piglet neonatal diarrhoea is an important issue in modern pig production and is linked to increased mortality and poor growth rates, affecting long-term pig health, increasing use of medication and cost of production. Saccharomyces cerevisiae var. boulardii (SB) is a probiotic yeast with documented clinical efficacy in the prevention and treatment of diarrhoeal diseases in humans. The objectives of the current study were to evaluate the effect of SB on occurrence and severity of neonatal diarrhoea in piglets, mortality and growth rate. Forty-six litters (606 piglets) were randomly allocated to a control or SB treatment (n=23 per treatment). Within 24 h of farrowing, piglets assigned to the SB treatment received a single oral dose of a paste containing 3.3×109 CFU of SB CNCM I-1079. Piglets from the control litters received a placebo paste. Piglet weight, mortality and diarrhoea were recorded up to day 7 of age. It was shown that numbers of diarrhoea days were significantly correlated with increased mortality rate and reduced weight gain (P<0.05). SB treatment had no effect on growth or mortality in diarrhoeic litters. However, SB-supplemented litters had significantly lower faecal scores, indicating firmer faeces (P<0.01) and fewer numbers of diarrhoeic days (P<0.01) during the 1st week of life. Reduction in the number of diarrhoeic litters compared with the control group was observed following the probiotic administration (P<0.05). These results highlight the detrimental effects of neonatal diarrhoea on pre-weaning performance and suggest that SB, by reducing diarrhoea duration and severity, has the potential of improving enteric health in the early stages of life in pigs.

Diarrhea outbreaks in suckling piglets due to rotavirus group C single and mixed (rotavirus groups A and B) infections

Porcine group A rotavirus (PoRVA) is a major cause of neonatal diarrhea in suckling and recently weaned piglets worldwide. The involvement of non-group A rotavirus in cases of neonatal diarrhea in piglets are sporadic. In Brazil there are no reports of the porcine rotavirus group C (PoRVC) as etiologic agent of the diarrhea outbreaks in piglets. The aim of this study was to describe the identification of rotavirus group C in single and in mixed infection with rotavirus groups A and B in three neonatal diarrhea outbreaks in suckling (&lt;21-day-old) piglets, with 70% to 80% and 20% to 25% of morbidity and lethality rates, respectively, in three pig herds located in the state of Santa Catarina, Brazil. The diagnosis of PoRV in the diarrheic fecal samples was performed using polyacrylamide gel electrophoresis (PAGE) to identify the presence of porcine rotavirus groups A, B (PoRVB), and C, and by RT-PCR (PoRVA and PoRVC) and semi-nested (SN)-PCR (PoRVB) to partially amplify the VP4 (VP8*)-VP7, NSP2, and VP6 genes of PoRVA, PoRVB, and PoRVC, respectively. One RT-PCR (PoRVA and PoRVC) and SN-PCR (PoRVB) product of each group of rotavirus of each diarrhea outbreak was submitted to nucleotide (nt) sequence analysis. Based on the PAGE technique, 4 (25%) and 1 (6.25%) of the 16 diarrheic fecal samples evaluated in the first outbreak presented PoRVA and PoRVC electropherotype, respectively, and 11 (68.75%) were negative. In the second outbreak, 3 (42.85%) of the 7 fecal samples evaluated presented PoRVA electropherotype, and in 3 (42.85%) and in 1 (14.3%) fecal samples were detected inconclusive and negative results, respectively. Three (30%) of the 10 fecal samples of the third outbreak presented PoRVC electropherotype; 5 (50%) and 2 (20%) samples showed negative and inconclusive results, respectively. Based on the RT-PCR and SN-PCR assays in the first neonatal diarrhea outbreak, PoRVC was detected in 13 (81.2%) of the 16 diarrheic fecal samples evaluated. PoRVC single infection was identified in 4 (25%) of these samples and mixed infections with PoRVA and PoRVB in 9 (56.2%) fecal samples. All of the seven diarrheic fecal samples evaluated from the second neonatal diarrhea outbreak were positive for PoRVC, whereas its mixed infection with other PoRV groups was detected in 4 (57.2%) samples. In the third outbreak, PoRVC in single infection was detected in all of the 10 diarrheic fecal samples analyzed. In the nt sequence analysis, the PoRVA strains of the first and second outbreaks demonstrated higher nt identity with G4P[6] and G9P[23] genotypes, respectively. The PoRVB strains (first and second outbreaks) and the PoRVC strains (first, second, and third outbreaks) showed higher nt identity and clustered in the phylogenetic tree with PoRVB and PoRVC strains that belong to the N4 and I1 genotypes, respectively. This is the first description in Brazil of the involvement of PoRVC in the etiology of diarrhea outbreaks in suckling piglets. The results of this study demonstrated that PoRVC, in both single and mixed infections, is an important enteropathogen involved in neonatal diarrhea outbreaks in piglets and that the use of more sensitive diagnostic techniques allows the identification of mixed infections involving two or even three groups of PoRV, which may be more common than previously reported.

Beta2 toxin is not involved in in vitro cell cytotoxicity caused by human and porcine cpb2-harbouring Clostridium perfringens

Clostridium perfringens is a common cause of intestinal disease in animals and humans. Its pathogenicity is attributed to the toxins it can produce, including the beta2 toxin. The presence of cpb2, the gene encoding the beta2 toxin, has been associated with diarrhoea in neonatal piglets and humans. However, the exact role of the beta2 toxin in the development of diarrhoea is still unknown. In this study we investigated the level of cytotoxicity to porcine IPI-21 and human Caco-2 cell-lines caused by porcine and human cpb2-harbouring C. perfringens and the significance of the beta2 toxin for the induction of cell cytotoxicity. Supernatants of porcine cpb2-harbouring C. perfringens strains were cytotoxic to both cell lines. Cell cytotoxicity caused by supernatant of human cpb2-harbouring C. perfringens strains was variable among strains. However, removal of the beta2 toxin by anti-beta2 toxin antibodies or degradation of the beta2 toxin by trypsin did not reduce the cytotoxic effect of any of the supernatants. These data suggest that beta2 toxin does not play a role in the development of cell cytotoxicity in in vitro experiments. In vivo studies are necessary to definitely define the role of beta2 toxin in the development of cell cytotoxicity and subsequent diarrhoea.

Dietary N-Carbamylglutamate Supplementation Boosts Intestinal Mucosal Immunity in Escherichia coli Challenged Piglets

N-carbamylglutamate (NCG) has been shown to enhance performance in neonatal piglets. However, few studies have demonstrated the effect of NCG on the intestinal mucosal barrier. This study was conducted to determine the effects of dietary NCG supplementation on intestinal mucosal immunity in neonatal piglets after an Escherichia coli (E. coli) challenge. New-born piglets (4 d old) were assigned randomly to one of four treatments (n = 7), including (I) sham challenge, (II) sham challenge +50 mg/kg NCG, (III) E. coli challenge, and (IV) E. coli challenge +50 mg/kg NCG. On d 8, pigs in the E. coli challenge groups (III and IV) were orally challenged with 5 mL of E. coli K88 (108 CFU/mL), whereas pigs in the sham challenge groups (I and II) were orally dosed with an equal volume of water. On d 13, all piglets were sacrificed, and samples were collected and examined. The results show that average daily gain in the E. coli challenged piglets (III and IV) was decreased (PE.coli<0.05). However, it tended to be higher in the NCG treated piglets (II and IV). Ileum secretory IgA, as well as IFN-γ, IL-2, IL-4 and IL-10 in ileal homogenates, were increased in E. coli challenged piglets (III and IV). Similarly, ileum SIgA and IL-10 levels, and CD4+ percentage in NCG treated piglets (II and IV) were higher than no-NCG treated piglets (PNCG<0.05). However, the IL-2 level was only decreased in the piglets of E. coli challenge + NCG group (IV) compared with E. coli challenge group (III) (P<0.05). No change in the IL-2 level of the sham challenged piglets (III) was observed. In conclusion, dietary NCG supplementation has some beneficial effects on intestinal mucosal immunity in E. coli challenged piglets, which might be associated with stimulated lymphocyte proliferation and cytokine synthesis. Our findings have an important implication that NCG may be used to reduce diarrhea in neonatal piglets.

The epidemiology of Clostridium perfringens type A on Ontario swine farms, with special reference to cpb2-positive isolates

BackgroundThere is poor understanding of most aspects of Clostridium perfringens type A as a possible cause of neonatal diarrhea in piglets, and the prevalence and types of C. perfringens present on Ontario swine farms is unknown. To study the prevalence of fecal C. perfringens and selected toxin genes, 48 Ontario swine farms were visited between August 2010 and May 2011, and 354 fecal samples were collected from suckling pigs, lactating sows, weanling pigs, grower-finisher pigs, and gestating sows, as well as from manure pits. The fecal samples were cultured quantitatively, and toxin genes were detected by real-time multiplex polymerase chain reaction (PCR).ResultsIn mixed multivariable linear analysis, log10C. perfringens in fecal samples from suckling pigs were higher than that of weanling pigs, grower-finisher pigs, and manure pit samples (P <0.05). In mixed multivariable logistic analysis, the C. perfringens isolates recovered from lactating sows (OR = 0.069, P <0.001), gestating sows (OR = 0.020, P <0.001), grower-finishers (OR = 0.017, P <0.001), and manure pits (OR = 0.11, P <0.001) were less likely to be positive for the consensus beta2 toxin gene cpb2 compared to the isolates from suckling pigs. The prevalence of cpb2 in the isolates recovered from weanlings did not differ significantly from suckling pigs. C. perfringens isolates that were positive for cpb2 were more likely to carry the atypical cpb2 gene (atyp-cpb2) (OR = 19, P <0.001) compared to isolates that were negative for cpb2. Multivariable analysis did not identify farm factors affecting the presence of consensus cpb2 and atyp-cpb2 genes.ConclusionsThis study provides baseline data on the prevalence of C. perfringens and associated toxin genes in healthy pigs at different stages of production on Ontario swine farms. The study suggests that if C. perfringens type A are involved in neonatal enteritis, there may be strains with specific characteristics that cannot be identified by the existing genotyping system.

Matched case-control study evaluating the frequency of the main agents associated with neonatal diarrhea in piglets

A case-control study was carried out in litters of 1 to 7-day-old piglets to identify the main infectious agents involved with neonatal diarrhea in pigs. Fecal samples (n=276) from piglets were collected on pig farms in the State of Rio Grande do Sul, Brazil, from May to September 2007. Litters with diarrhea were considered cases (n=129) and normal litters (n=147) controls. The samples were examined by latex agglutination test, PAGE, conventional isolating techniques, ELISA, PCR, and microscopic methods in order to detect rotavirus, bacterial pathogens (Escherichia coli, Clostridium perfringens type A and C, and Clostridium difficile), and parasites (Coccidian and Cryptosporidium spp.). Outbreaks of diarrhea were not observed during sampling. At least one agent was detected in fecal samples on 25 out of 28 farms (89.3%) and in 16 farms (57.1%) more than one agent was found. The main agents diagnosed were Coccidia (42.86%) and rotavirus (39.29%). The main agents identified in litters with diarrhea were Clostridium difficile (10.6%), Clostridium perfringens type A (8.8%) and rotavirus (7.5%); in control litters, Clostridium difficile (16.6%) and Coccidian (8.5%). Beta hemolytic Escherichia coli and Clostridium perfringens type C were not detected. When compared with controls, no agent was significantly associated with diarrhea in case litters. These findings stress the need for caution in the interpretation of laboratorial diagnosis of mild diarrhea in neonatal pigs, as the sole detection of an agent does not necessarily indicate that it is the cause of the problem.

AVALIAÇÃO DE ÓLEOS ESSENCIAIS DE ORÉGANO E ALECRIM NA PREVENÇÃO DA DIARREIA NEONATAL EM LEITÕES

RESUMO Com objetivo de verificar o efeito preventivo da emulsão líquida de uma mescla de óleos essenciais de orégano (Origanum vulgare) e alecrim (Rosmarinus officinalis) no controle da diarreia neonatal em leitões lactantes foi realizado um experimento composto por dois tratamentos simples em um delineamento experimental inteiramente casualizado. Leitões de mesma leitegada, nascidos de fêmeas com ordens de partos variadas, foram divididos em 2 grupos de leitões sem manifestação clínica de diarreia neonatal. O primeiro grupo com 661 leitões recebeu o tratamento controle (1 mL kg-1 solução fisiológica) e o segundo grupo com 672 animais recebeu óleos essenciais de orégano (60% de carvacrol e 15% timol) e 2% de óleo essencial de alecrim, sob a forma de emulsão líquida a 5% na dose de 1 mL kg-1.. Todos os tratamentos foram administrados via oral no primeiro e terceiro dias de vida do suíno, sendo que, para o parâmetro índice de prevenção e causas de mortalidade dos leitões, cada animal representou uma repetição. Não houve diferença significativa entre os tratamentos quanto ao índice de prevenção da diarreia neonatal em leitões. Os resultados permitiram concluir que a mescla de óleos essenciais à base de orégano e alecrim não previne as diarreias neonatais dos leitões.

Homeopathy as replacement to antibiotics in the case of Escherichia coli diarrhoea in neonatal piglets

The use of antibiotics in the livestock sector is increasing to such an extent that it threatens negative consequences for human health, animal health and the environment. Homeopathy might be an alternative to antibiotics. It has therefore been tested in a randomised placebo-controlled trial to prevent Escherichia coli diarrhoea in neonatal piglets. On a commercial pig farm 52 sows of different parities, in their last month of gestation, were treated twice a week with either the homeopathic agent Coli 30K or placebo. The 525 piglets born from these sows were scored for occurrence and duration of diarrhoea. Piglets of the homeopathic treated group had significantly less E. coli diarrhoea than piglets in the placebo group (P<.0001). Especially piglets from first parity sows gave a good response to treatment with Coli 30K. The diarrhoea seemed to be less severe in the homeopathically treated litters, there was less transmission and duration appeared shorter.

Isolation and identification of AIDA-I receptors in porcine intestinal mucus

Porcine AIDA-I positive Escherichia coli causes diarrhea in neonatal piglets and AIDA-I adhesin is an important virulence factor involved in intestinal colonization with biofilm formation. This biofilm consists of AIDA-I + E. coli bacteria stratified within mucus layers covering the intestinal mucosa. Based on the intimate interaction between AIDA-I + E. coli and mucus within the intestinal biofilm, we hypothesized that porcine intestinal mucus contains receptor(s) for AIDA-I adhesin. Since porcine AIDA-I receptors have not been identified, we employed affinity chromatography and in vitro adhesion assays to investigate AIDA-I binding proteins in porcine intestinal mucus that might serve as receptors for attachment of AIDA-I positive E. coli. We demonstrated that porcine mucus contains 65 and 120 kDa proteins (p65 and p120) that bind with high affinity to purified AIDA-I adhesin and that AIDA-I positive E. coli binds to these proteins with higher affinity than do AIDA-I negative mutant. The identity of p65 was not determined based on LC–MS/MS data, whereas p120 was matched to two nuclear proteins (namely, DNA damage binding protein and splicing factor 3b) and one cytoplasmic protein, which is an IgG Fc binding protein. Based on similar amino acid homology, molecular weight, structural similarity to mucin and reported evidence of being secreted by goblet cells into the intestinal lumen, we think that the IgG Fc binding protein is most likely candidate to serve as a potential receptor in intestinal mucus for AIDA-I adhesin.