Distribution of Salmonella serovars and phage types on 80 Ontario swine farms in 2004

The objective of this study was to describe the distribution of Salmonella spp. on Ontario grower-finisher pig farms. Eighty swine farms were visited from January through July 2004. On each farm, fecal samples were collected from 5 pens, 2 rectal samples and 1 pooled sample from fresh manure on the floor per pen. Salmonella was isolated from 91 (11%) of the 800 rectal samples and 73 (18%) of the 397 pooled samples. Overall, Salmonella was recovered from 37 (46%) of the 80 farms. On each positive farm, Salmonella was cultured from 1 to 7 pigs or 1 to 5 pens. Of the 37 farms, 18, 13, 5, and 1 yielded 1, 2, 3, and 4 serovars, respectively. The most common serovars were S. Typhimurium var. Copenhagen, S. Infantis, S. Typhimurium, S. Derby, S. Agona, S. Havana, and S. enterica subsp. I:Rough-O. The 3 most frequent phage types were PT 104, PT 104a, and PT 104b. There was a statistically fair agreement between samples collected directly from pigs and pooled pen samples in determining the Salmonella status at the pen and farm level (kappa = 0.6, P < 0.0001). However, in 62 pens, Salmonella status, serovars, or phage types differed between the pig and pooled pen samples. The distribution of Salmonella on the swine farms in this study indicates that, in developing an intervention strategy, priority should be given to farms positive for S. Typhimurium var. Copenhagen. Also, the variation in Salmonella status between pig and pooled pen samples deserves consideration in a sampling strategy.     

Prevalence of rotavirus A infection and the detection of type G3P[11] strain in ruminants in Yobe state,Nigeria

Rotaviruses have a worldwide distribution and the infection is associated with diarrhea in young of ruminants as well as children. However, limited data exist on its prevalence and types in Yobe state, Nigeria. Detection of rotavirus A and types in ruminant population in Yobe state was the aim of the study. A total of 470 diarrheic fecal samples were collected and tested for rotavirus and types using serology and molecular techniques respectively. A prevalence rate of 2.98% (14/470) was found in the three species with specific rates of 2.9% (6/202), 3.8% (6/158), and 1.8% (2/110) in goat, sheep, and cattle respectively. The prevalence rates of 3.6% (12/331), 1.2% (1/84), and 1.8% (1/55) were for those aged < 1–3, 4–6, and 7–9 months old, respectively, while 4.9% (9/185) and 1.7% (5/285) were in males and females respectively. Rotavirus genes VP7 and VP4 were detected in 2 (14.3%) out of the 14 ELISA-positive samples while deduced amino acid sequences of the major variable regions revealed the genes to belong to types G3P[11] strain. Significant association was found between the infection and sex (P < 0.05) unlike in the species and age groups of the ruminants. The circulation of rotavirus virus in ruminants and type G3P[11] in cattle has been confirmed in the study. Hence, there is a need for continuous surveillance, awareness campaign, and assessment of the economic losses and public health implications of rotavirus infection in Nigeria.

     

Epidemiology and antibacterial susceptibility patterns of bloodstream infections, 2001-2004: an experience with BACTEC 9240 in Southern Iran

This study was conducted to determine the prevalence of bacteria recovered from bloodstream samples by Bactec 9240 at our hospital wards and to evaluate their antibacterial susceptibility patterns. During January 2001 through December 2004, 9407 referred blood samples in Bactec bottles from admitted patients at three main wards, neonates, pediatrics and adults at Nemazee Hospital, affiliated to Shiraz University of Medical Sciences in Shiraz were processed. Positive cultures were purified and identified according to standard methods. Sensitivity of bacteria to different antibiotics was determined by Kirby-Bauer disk diffusion method. Staphylococcus aureus 132(25%), Escherichia coli 64(12.1%) and Pseudomonas aeruginosa 52(9.8%) were the most pathogenic bacteria which were recovered from the blood samples. Pathogenic microorganisms were isolated from blood samples of 305 (57.8%) at pediatrics, from 181 (34.2%) at adults and from 42 (8%) at neonates wards. The highest antibiotics activities against gram positive isolates observed for vancomycin (98.4%), chloramphenicol (86.4%) and ciprofloxacin (77.4%), while in gram negative bacteria imipenem (96.1%), ciprofloxacin (83%) and amikacin (77.9%), were effective antibiotics. Frequency of isolated bacteria at pediatrics compared to adults and neonates wards were approximately two and seven folds high, respectively which indicates special attention should be paid to pediatrics patients both in prevention and treatment aspects. Vancomycin and imipenim are the effective antibiotics and could cover majority of gram positive and negative bacteria. Therefore, combined administrations of these antibiotics seems mandatory for empirical therapy.     

Molecular epidemiology and antimicrobial resistance of Salmonella typhimurium DTI04 on Ontario swine farms.

This study was conducted to examine antimicrobial resistances, plasmid profiles, and pulsed-field gel electrophoresis patterns of 80 Salmonella Typhimurium (including var. Copenhagen) DT104 strains (including DT104a and DT104b) recovered from pig and environmental fecal samples on 17 swine farms in Ontario. No resistance was observed to amoxicillin/clavulanic acid, apramycin, carbadox, cephalothin, ceftriaxone, ceftiofur, cefoxitin, ciprofloxacin, nalidixic acid, trimethoprim, and tobramycin. However, the isolates exhibited resistance against 4 to 10 antimicrobials with the most frequent resistance being to sulfonamides (Su), ampicillin (A), streptomycin (S), spectinomycin (Sp), chloramphenicol (C), tetracycline (T), and florfenicol (F). Thirteen distinct resistance patterns were determined but 88% of isolates shared the typical resistance pattern "ACSpSSuT." Twelve different plasmid profiles were observed; the 62 MDa virulence-associated plasmid was detected in 95% of the isolates. The 2.1 MDa plasmid was the second most frequent one, which was harbored by 65% isolates. The isolates were classified into 23 distinct genotypes by PFGE-SpeI + BlnI when difference in at least one fragment was defined as a distinct genotype. In total, 39 distinct "types" were observed when defining a "type" based on the combination of antimicrobial resistance, plasmid pattern, and PFGE-SpeI + BlnI for each isolate. The highest diversity was 0.96 (95% CI: 0.92, 0.96) for the "type" described above followed by 0.92 (95% CI: 0.88, 0.93) for PFGE-SpeI + BlnI. The diversity of DT104 isolates indicates there might be multiple sources for this microorganism on swine farms. This knowledge might be used to track these sources, as well as to study the extent of human salmonellosis attributed to pork compared to food products derived from other food-producing animals.     

A clinical field trial to evaluate the efficacy of vaccination in controlling Salmonella infection and the association of Salmonella-shedding and weight gain in pigs

A clinical field trial was performed to determine the effectiveness of an autogenous Salmonella Typhimurium bacterin compared with a commercial live S. Choleraesuis vaccine in pigs. The association between Salmonella shedding and weight gain was also investigated. Nine cohorts of weaned pigs, (330 to 350 pigs per cohort), were randomly assigned to 1 of 3 treatment groups (injection with S. Typhimurium bacterin, vaccination via water with S. Choleraesuis vaccine, or a control group receiving no vaccine). In each cohort, the average daily gain was calculated for a selected pen throughout the production stage. Pen (pooled) fecal samples were collected bi-weekly and cultured. The odds of Salmonella shedding in both vaccinated groups was higher than in the control group (P < 0.05). The prevalence of Salmonella shedding declined overall as pigs aged (P = 0.04). However, the control pigs showed the smallest decrease in Salmonella shedding over the entire production stage, while prevalence of Salmonella shedding in the vaccinated groups decreased twice as much as the control group over the entire production stage. Salmonella Typhimurium var. Copenhagen DT104, S. Cerro, and S. Agona, which had been isolated on the study farm previously, were recovered from pigs in this study. Shedding of S. Typhimurium var. Copenhagen decreased over time in both vaccine treatment groups. On the other hand, S. Cerro shedding rate was lower in the control pigs compared with vaccinated pigs and S. Agona could be recovered only from the samples collected from S. Choleraesuis vaccinated pigs. The pigs from pens with a higher Salmonella recovery rate experienced slower growth compared with pigs from pens where Salmonella was not isolated. This latter finding indicates that there might be an economic incentive for producers to try to control endemic salmonellosis if effective programs could be developed.     

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.     

Use of dye industry effluent for irrigation in Amaranthus cruentus L. production: effect on growth,root morphology,heavy metal accumulation,and the safety concerns

This study characterized three local dye effluents from major tie and dye industries at Asero, Itoku, and Kemta in Ogun State, Nigeria, and evaluated their effect on growth and heavy metal accumulation in pot-grown amaranth. The effluents were diluted with water at 0%, 25%, 50%, and 100% and replicated three times. Physicochemical properties and heavy metal content of effluent dilutions were analyzed. Results indicated that irrespective of the source, effluents, and dilutions, effluent properties exceeded permissible limits by World Health Organization/Food and Agricultural Organization/Federal Environmental Protection Agency for irrigation. The SO₄^2?, PO₄^3?, NO₃^?, Fe, and Cr contents of effluent dilutions were higher than the maximum limits recommended. The order of toxicity of dye effluents was Asero > Itoku > Kemta. Effluent concentrations at 50% and 100% damaged amaranth root hairs, caused the death of some amaranth, significantly reduced number of leaves, height, fresh and dry root, and shoot weight, but increased stem girth and heavy metal accumulation in plant parts. Mg, Cu, and Zn were more concentrated in the shoots. Due to the high metal content, consumption of this vegetable poses a potential health risk. Strict law enforcement against the use of the effluent for irrigation should be made to prevent environmental hazard.     

Prevalence of Salmonella spp. on Canadian pig farms using liquid or dry-feeding

The objective of this study was to determine whether the shedding and antibody titre to Salmonella was lower for pig herds provided liquid-feed compared to those on traditional dry rations. Twenty liquid-feeding farms and 61 dry-feeding farms were selected. The amount of antibodies to Salmonella in sera from 15 finisher pigs on each of 80 Ontario swine farms was analyzed by means of enzyme-linked immunosorbent assay (ELISA). In addition, the presence of Salmonella on the 20 liquid-feeding farms and 21 of the dry-feeding farms was assessed by culture of 15 fecal samples taken directly from finisher pigs and five pooled pen-fecal samples at each farm.

A cut-off of OD% 10 was used. The Salmonella sero-prevalence differed between the two groups of farms. At least one pig tested sero-positive on 98% of the dry-feeding farms and 84% of the liquid-feeding farms (P < 0.05). A multi-variable mixed linear regression model with the farm as a random variable and farm factors as the fixed effects was fitted. Crude optical density (OD) of the individual pig was considered as the continuous dependent variable. Dry-feeding and antimicrobial daily usage was associated with crude OD (P < 0.05). In addition, crude OD increased with increasing herd size (P < 0.05).

Salmonella was isolated from 25 out of 420 fecal samples (6%) from dry-feeding farms compared to three out of 400 samples (0.8%) from liquid-feeding farms. Eight of the dry-feeding farms (38%) tested positive compared to only three of the liquid-feeding farms (15%). Salmonella was also recovered from the pen environment on five dry-feeding farms but were not isolated from the facilities using liquid-feeding. Salmonella Typhimurium was isolated from four farms in the dry-feed group and on one farm with liquid-feeding. The one S. Typhimurium isolate from the liquid-feeding farm exhibited no antimicrobial resistance, but those from dry-feeding farms were resistant to four or more antimicrobial agents. The results of the logistic regression, with farm as a random effect showed that dry-feeding [OR = 2.7 (1.1–15.1)] and continuous flow system [OR = 2.3 (1.2–12.7)] increased risk of finding Salmonella in the individual pig.

These findings indicate that liquid-feeding and all-in all-out management of the grower–finisher barns can reduce the Salmonella prevalence.     

A retrospective study on the etiological diagnoses of diarrhea in neonatal piglets in Ontario,Canada, between 2001 and 2010

Laboratory surveillance data from the Animal Health Laboratory, University of Guelph, on the etiological diagnoses of neonatal diarrhea in piglets were analyzed to determine the relative importance and trends of different enteric pathogens in Ontario. A total of 237 cases, including live and dead 1- to 7-day-old piglets, were submitted for diagnosis of gastrointestinal illness between 2001 and 2010. The combined frequencies for cases of gastrointestinal illness involving Escherichia coli, Clostridium perfringens type A, rotavirus, and Clostridium difficile, either as single pathogens or a complex of pathogens, accounted for 56% of the total cases. In a total of 33% of cases of gastrointestinal illness, an etiological agent was not identified. The frequency of cases diagnosed with enterotoxigenic E. coli was decreased from 2007. Cases submitted in 2010 were more likely to be diagnosed with C. perfringens type A compared to cases submitted in 2002 to 2007 (P < 0.05). There was a significant trend for cases submitted in the winter to be diagnosed with C. perfringens type A, enterotoxigenic E. coli, rotavirus, and Cystoisospora suis (formerly Isospora suis) (P < 0.05). Enterotoxigenic E. coli was less likely diagnosed if C. difficile, C. perfringens, or rotavirus were detected (P < 0.05). Younger piglets were more likely to be diagnosed with C. perfringens type A (P < 0.05) and C. difficile (P < 0.05) than older piglets. This study shows that E. coli, C. perfringens type A, rotavirus, and C. difficile are enteric pathogens of concern for Ontario swine farrowing operations and further research is required to understand the reasons for the cases that are not diagnosed.     

Vitrification of mouse preantral follicles versus slow freezing: Morphological and apoptosis evaluation

The aim of this study was evaluation of survivability, maturation rate and apoptotic gene expression of preantral follicles after vitrification and slow freezing technique. Normal mouse preantral follicles were randomly divided into three experimental groups. In the control group, follicles were cultured immediately; in the vitrification and slow freezing groups, follicles were cultured after vitrification‐warming and slow freezing‐thawing procedures. Follicular viability was assessed by using 0.4% trypan blue, and molecular evaluation of messenger RNA levels of apoptosis‐related genes was performed by the semi‐quantitative RT‐PCR method after 3 h of culture. Oocyte maturation rates were also evaluated on day 14 of culture. Survival and maturation rate in the slow freezing group were significantly lower than those in control and vitrification groups (P ≤ 0.05). Although there was no difference in Survivin expression among the three experimental groups, Bcl‐2 expression was significantly lower in the slow freezing group compared to the other groups (P ≤ 0.05). The expression of Bax, P53, Fas and Bax/Bcl‐2 ratio in the slow freezing group was significantly higher than control and vitrification groups (P ≤ 0.05). Preantral follicle vitrification seems to be better than slow freezing as seen in the survival, maturation and expression rates of apoptotic gene variants.