Non-diarrheic Dogs Research Articles

PREVALENCE OF TOXOCARA CANIS AND ASSESSMENT OF KNOWLEDGE, ATTITUDE, AND PRACTICES OF THE COMMUNITY IN BISHOFTU TOWN, CENTRAL ETHIOPIA

A cross-sectional study was conducted in order to determine the prevalence of Toxocara canis, potential risk factors, and knowledge, attitude, and practices in Bishoftu, central Ethiopia. Fecal samples were taken from 300 randomly selected dogs of different sex, age group, breeds, and diarrheal status. The samples were processed using a flotation technique to detect the eggs of Toxocara canis. The current result revealed an overall prevalence of 32.33% (95% CI: 27.07 - 37.95) of Toxocara canis. The prevalence of the disease was 30.30% (95% CI: 22.61 - 38.90), 31.06% (95% CI: 23.30 - 39.70), and 44.44% (95% CI: 27.94 - 61.90) in young, adult and older dogs, respectively. The prevalence of Toxocara canis was 32.43% (95% CI: 26.77 - 38.50) and 31.71% (95% CI: 18.08 - 48.09) in female and male dogs, respectively. The present findings indicated that there were statistically significant links between the prevalence of Toxocara canis regarding dog breeds and diarrheal status (p < 0.05). The prevalence was higher in German Shepherd breed dogs (56.52%; 95% CI: 34.49 - 76.81) than in local dog breeds (30.32%; 95% CI: 24.97 - 36.11). The prevalence of the disease was higher in diarrheic dogs (46.48%; 95% CI: 34.55 - 58.71) than in non-diarrheic dogs (27.95%; 95% CI: 22.24 - 34.24). The knowledge, attitude, and practice assessment in the present study indicated poor dog management practices and insufficient public awareness. Most dog owners (56.7%; 95% CI: 50.85 - 62.35) did not deworm their dogs regularly, while 90.67% (95% CI: 86.79 - 93.71) fed dogs raw animal products. The present finding indicated that most respondents were not aware of zoonotic canine parasites (83.7%; 95% CI: 81.18 - 89.43) and toxocariasis (80.7%; 95% CI: 75.74 - 84.98). This study demonstrates a higher prevalence of Toxocara canis and lower public awareness about the disease. Therefore, this study recommends strategic deworming of dogs, better dog management, and increased public awareness in order to control the impact of the disease on animal and human health.

Giardiasis and diarrhea in dogs: Does the microbiome matter?

Giardia duodenalis (Gd) causes intestinal parasitosis. The involvement of the intestinal microbiome in determining the infection's clinical phenotype is unknown. Investigate the fecal microbiome features in dogs with giardiasis. Cross-sectional study, including fecal samples of kenneled dogs with Gd diagnosed by fecal Giardia antigen dot ELISA. The fecal microbial compositional characteristics and dysbiosis index (DI) were compared between diarrheic and nondiarrheic dogs. Fecal samples of 38 Gd-infected dogs (diarrheic, 21; nondiarrheic, 17) were included. No differences were found in Faith's phylogenic diversity and beta diversity (weighted UniFrac distances) and in specific taxa abundances at the phylum, genus, and species levels, as well as in alpha and beta diversities between diarrheic and nondiarrheic dogs, and also when divided by sex or age. Among diarrheic dogs, alpha diversity was higher in males than in females (pairwise Kruskal-Wallis, q = 0.01). Among males, fecal abundances of the genus Clostridium (W = 19) and Clostridium spiroforme species (W = 33) were higher in diarrheic compared to nondiarrheic dogs. In diarrheic dog fecal samples, Proteobacteria were more prevalent (W = 1), whereas Verrucomicrobia were less prevalent in dogs <1 year of age than in older dogs. The fecal sample DI of 19 diarrheic and 19 nondiarrheic dogs was similar (median, -0.2; range, -4.3 to 4.5 and median, -1.0; range, -4.3 to 5.8, respectively). The fecal microbial composition of symptomatic and asymptomatic dogs with giardiasis is similar. Based on fecal DI, giardiasis is not characterized by prominent dysbiosis. Other host and parasite characteristics might determine the severity of giardiasis in dogs.

DETECTION OF SOME VIRULENCE, ANTIBACTERIAL RESISTANCE GENES FOR SALMONELLA ISOLATED FROM DOGS IN BAGHDAD CITY

Two hundred and fifty (250) samples from both diarrheic and non-diarrheic dogs in both sex male and female multi breeds ( German shepherd and malinois ) in Academy for training police and military dogs for searching about drugs , explosives ,local dogs and pet dogs from different clinical cases in Baghdad city were tested. All samples are obtained in aseptically rectal swabs and transporting to the microbiology lab, for culturing and identification for salmonella species. The study gave 16 (6.4%) positive Salmonella. Virulent gene study gave us 16(6.4%) invA gene, invF 13(5.2%) and sit C 16 (6.4%).Antibiotic sensitivity test According to Bauer-Kirby method were done and gave 16 (6.4%) isolates resistance for ciprofloxacin which is the most clinical antibiotic drug of choice against Salmonella infection . Detection of gyrA gene resistance gene for ciprofloxacin using Polymerase chain reaction (PCR) gave 16 (6.4%) for all positive isolates.

Prevalence of resistance and virulence genes in Escherichia coli isolates from diarrheic dogs

The current study was designed to determine the prevalence of antimicrobial resistance and virulence genes in Escherichia coli isolated from canine diarrheal cases. Fecal samples were collected from 77 dogs with clinical diarrhea and 10 non-diarrheic dogs. Breed, age and sex, and clinical manifestations were recorded. Samples were plated on MacConkey and EMB agars, followed by standard isolation procedures and identification of E. coli. An antimicrobial susceptibility test using the disc diffusion method was performed against Ampicillin, Tetracycline, Trimethoprim, Gentamicin, and Streptomycin. PCR was used to determine if the isolates carry virulence intimin adherence protein (eaeA) and antimicrobial resistance genes. Body temperature, respiratory, and heart rates in dogs with diarrhea were significantly higher than in non-diarrheic dogs. PCR detected the eaeA virulence gene in 44(69.8%) of 63 isolates from diarrheic dogs. All isolates were resistant to ampicillin, and 55(87.3%), 47(74.6%), 29(46.0%), and 19(30.2%) of these were resistant to tetracycline, trimethoprim, gentamicin, and streptomycin, respectively. The frequency of antimicrobial resistance genes in the 63 isolates was 81.0, 52.4, 41.3, 33.3, 23.8 and 9.5% for CITM, tet (B), dfrA1, aac (3)-IV, aadA1 and tet (A), respectively. Overall, 6(9.4%), 16(25.4%), and 41(65.1%) were positive for one, two, three, or four resistance genes, respectively. In conclusion, the high prevalence of virulence (69.8%) and resistance 9.5-81.0% genes in E. coli isolates could be responsible for the diarrhea episodes, which may have posed therapeutic implications in affected dogs.

Carnivore chaphamaparvovirus-1 (CaChPV-1) infection in diarrheic dogs reveals viral endotheliotropism in intestine and lung

Background Carnivore chaphamaparvovirus-1 (CaChPV-1) is a parvovirus identified in dogs and association of infection with diarrhea is controversial. Information on whether tissue tropism persists is lacking. Objectives To determine the disease association of CaChPV-1 in dogs with diarrhea and to investigate viral tropism and genetic diversity. Animals and methods CaChPV-1 infection was investigated in five recently deceased puppies and designed a retrospective study to determine whether the presence of CaChPV-1 is associated with diarrhea. The retrospective study was conducted in 137 intestinal tissue samples and 168 fecal samples obtained from 305 dogs. CaChPV-1 tissue localization was determined using in situ hybridization, and CaChPV-1 complete genomes obtained from dead puppies and retrospective study were sequenced and analyzed. Results CaChPV-1 was detected in 6.56% (20/305) of tested dogs, including 14 diarrheic- and 6 non-diarrheic dogs, and was significant in puppies with diarrhea (p = 0.048). Among the CaChPV-1-positive diarrheic dogs, one sample was obtained from intestinal tissue and 13 samples were fecal samples. However, six CaChPV-1 positive non-diarrheic dogs were based on fecal samples but not on intestinal tissue. Within the age range, the presence of CaChPV-1 was significant in puppies (p < 0.00001) and was mainly localized in the stromal and endothelial cells of intestinal villi and pulmonary alveoli. Phylogenetic analysis indicated genetic diversity of CaChPV-1 Thai strains that were mostly clustered within the sequences found in China. Conclusions Although definitive pathogenesis of CaChPV-1 remains undetermined, this study provides evidence supporting that CaChPV-1 localizes in canine cells and could play a potential role as an enteric pathogen.

Detection and Genomic Characterization of Canine Circovirus in Iran

Simple SummaryDuring the last decade, canine circovirus (CaCV) has been repeatedly reported in domestic dogs with and without acute enteritis. Here we report the detection and full genome characterization of CaCV strains from non-diarrheic dogs in Iran. The results showed the circulation of the virus in this country, with Iranian strains segregating from the CaCVs detected in other countries.Canine circovirus (CaCV) is a single-stranded DNA virus that globally circulates in dogs and wild carnivores. Although the pathogenic potential of the virus has not been fully understood yet, CaCV has been suggested to exacerbate the clinical course of other canine viral infections but also to circulate in dogs without clinical signs. In this study, we carried out real-time PCR assays to detect enteric pathogens from 156 canine rectal swabs collected from dogs without enteritis in 3 different regions in Iran. A total of 14 samples tested positive for CaCV and full-length genome sequences were obtained from 6 of the detected strains. Sequence and phylogenetic analyses showed that, despite the distance between the different sample collection sites, all Iranian CaCV strains were closely related and formed a separate clade from extant CaCVs. The present study shows that CaCV is circulating in non-diarrheic dogs in Iran, thus highlighting the need for further epidemiological investigations in Iranian domestic and wild carnivores.

Isolation, antimicrobial susceptibility patterns, and risk factors assessment of non-typhoidal Salmonella from apparently healthy and diarrheic dogs

BackgroundDogs are one of the important asymptomatic carriers of antimicrobial resistant and potentially pathogenic strains of Salmonella. They can harbor large bacterial load in the intestines and mesenteric lymph nodes which can be shed in their feces with the possibility of transmission to humans. Therefore, a cross-sectional study was conducted with the objectives of estimating the prevalence of non-typhoidal Salmonella, assessing the risk factors for dog’s Salmonella carriage, and profiling the antimicrobial resistance pattern of Salmonella isolates among housed dogs in Harar town, Eastern Ethiopia. A total of 415 rectal swab samples were collected from randomly selected dogs. Samples were examined for non-typhoidal Salmonella using standard bacteriologic culture and biochemical tests. The disk diffusion method (Kirby-Bauer test) was employed to evaluate the isolates for their susceptibility against five antimicrobials.ResultsNon-typhoidal Salmonella were isolated from 26 (6.3%) of the rectal swab samples, with significantly higher occurrence in diarrheic (15.2%) than non-diarrheic (5.5%) dogs. The risk of Salmonella harboring was significantly higher in female dogs than in male dogs (OR = 2.5, p = 0.027). Dogs fecal shedding of Salmonella was relatively higher in households who used offal as a main feed type for their dogs (23.1%; 95% CI = 5–53.8) than those who used leftover food (10.1%; 95% CI = 5.7–16.1) and practiced mixed feeding system (17%; 95% CI = 7.6–30.8). Salmonella isolates showed higher resistance to ampicillin (41.7%), while all isolates were fully susceptible to gentamicin. Moreover, 58.3% of Salmonella isolates showed resistance to at least one of the tested antimicrobials. Majorities (72.7%) of the dog owners had no awareness on the risk of zoonotic salmonellosis from dog and all of the respondents use bare hand to clean dog kennel.ConclusionOur study reveals the importance of both diarrheic and apparently healthy housed dogs in the harboring and shedding of antimicrobial resistant non-typhoidal Salmonella. The risk of non-typhoidal Salmonella spread among pet owners is not negligible, especially in households who use offal as main feed type. Therefore, an integrated approach such as: proper dog handling practices; continuous evaluation of antimicrobial resistance; and rational use of antimicrobials in the field of veterinary sector are necessary to tackle the problem.

Prevalence and Molecular Identification of Giardiasis Isolates from Stray Dogs in Erbil Province, Kurdistan Region, Iraq

Giardia lamblia is the intestinal, flagellated protozoan parasite. It should make a species complex and comprises eight assemblages (A-H). In the current study, out of 153 examined samples, 16 (10.46%) and 36 (23.53%) specimens were positive by concentration and polymerase chain reaction (PCR) technique for giardiasis, respectively. The highest rate of infection was found in a rural area (14.38%) than in urban areas (9.15%). As well as, the infection rate in males (25/153) was higher compared to that of female dogs (11/153). Regarding fecal consistency, the highest rate (18.3%) of giardiasis was observed in diarrheic dogs, while the lowest rate of giardiasis (5.23%) was observed among non-diarrheic dogs. The PCR products were sequenced for 20 samples and further examined by sequence analysis, 16 isolates under the accession number (MN629930), independent of the host, exhibited G. lamblia GenBank ID: M36728, while in four samples under the accession number (MN629931), nucleotide substitutions generate polymorphism at position 542 and 561 (C → T) and (A → G) at position 684. The similarity between MN629931 and AY072723 genotype A2 was 99.9% which was one nucleotide substitution at position 542 (C → T). The sequencing of the PCR products recognized two assemblages in dogs suggested the possible role of dogs as the reservoir for human giardiasis in Erbil Province which is the first records in the Kurdistan Region, Iraq.

Antimicrobial resistance patterns of commensal Escherichia coli isolated from feces of non-diarrheic dogs in Grenada, West Indies.

Background and Aim:There is currently no published information on the prevalence and antimicrobial susceptibility patterns of commensal Escherichia coli in dogs of Grenada origin. Monitoring antimicrobial resistance helps in the empirical selection of antibiotics. This study determined the occurrence of E. coli including the O157:H7 serotype in feces of non-diarrheic dogs of Grenada origin and the antibiotic resistance pattern of the E. coli isolates.Materials and Methods:Fecal samples from 142 of the 144 (98.6%) dogs were culture positive for E. coli. Selection of up to three colonies from each of the 142 E. coli-positive samples yielded a total of 402 E. coli isolates, which were analyzed for the presence of non-sorbitol fermenting colonies, and O157-agglutination.Results:Of the 402 E. coli isolates, 30 (7.5%) were non-sorbitol fermenters. However, none of the 402 isolates gave a positive reaction (O157:H7) to the E. coli O157:H7 latex kit. Antimicrobial susceptibility tests against 12 antibiotics revealed low resistance rates to all the tested antibiotics except for tetracycline (Te) (23.4%), cephalothin (CF) (13.2%), and ampicillin (AM) (7.7%). Thirty-nine out of the 402 (9.7%), E. coli isolates were resistant to two or more antibiotics of different classes.Conclusion:This is the first report of isolation and antimicrobial susceptibilities of commensal E. coli from non-diarrheic dogs in Grenada. Some of the isolates (39/402 isolates, 9.7%) were resistant to multiple antibiotics. This study showed that presently, dogs in Grenada should not be considered a reservoir for the E. coli O157:H7 serotype and for multiple antibiotic-resistant E. coli strains. Among the 402 E. coli isolates, the resistance rate to drugs other than Te, CF, and AM was very low.

Prevalence, toxin gene profile, antibiotic resistance, and molecular characterization of Clostridium perfringens from diarrheic and non-diarrheic dogs in Korea.

Clostridium perfringens causes diarrhea and other diseases in animals and humans. We investigated the prevalence, toxin gene profiles, and antibiotic resistance of C. perfringens isolated from diarrheic dogs (DD) and non-diarrheic dogs (ND) in two animal hospitals in Seoul, Korea. Fecal samples were collected from clinically DD (n = 49) and ND (n = 34). C. perfringens was isolated from 31 of 49 DD (63.3%) and 21 of 34 ND dogs (61.8%). All C. perfringens strains were positive for the α toxin gene, but not for the β, ε, or ι toxin genes; therefore, all strains were identified as type A C. perfringens. All isolates were cpe-negative, whereas the β2 toxin gene was identified in 83.9% and 61.9% of isolates from DD and ND, respectively. Most isolates were susceptible to ampicillin (94%), chloramphenicol (92%), metronidazole (100%), moxifloxacin (96%), and imipenem (100%). However, 25.0% and 21.2% of isolates were resistant to tetracycline and clindamycin, respectively. Molecular subtyping of the isolated strains was performed by using pulsed-field gel electrophoresis. Fifty-two isolates were classified into 48 pulsotypes based on more than 90% similarity of banding patterns. No notable differences were observed among the isolates from DD and ND.

Detecção de genes de virulência e grupos filogenéticos de amostras Escherichia coli isoladas de cães no Brasil

ABSTRACT: This study identified the virulence genes, pathovars, and phylogenetic groups of Escherichia coli strains obtained from the feces of dogs with and without diarrhea. Virulence genes and phylogenetic group identification were studied using polymerase chain reaction. Thirty-seven E. coli isolates were positive for at least one virulence factor gene. Twenty-one (57.8%) of the positive isolates were isolated from diarrheal feces and sixteen (43.2%) were from the feces of non-diarrheic dogs. Enteropathogenic E. coli (EPEC) were the most frequently (62.2%) detected pathovar in dog feces and were mainly from phylogroup B1 and E. Necrotoxigenic E. coli were detected in 16.2% of the virulence-positive isolates and these contained the cytotoxic necrotizing factor 1 (cnf1) gene and were classified into phylogroups B2 and D. All E. coli strains were negative for the presence of enterotoxigenic E. coli (ETEC) enterotoxin genes, but four strains were positive for ETEC-related fimbriae 987P and F18. Two isolates were Shiga toxin-producing E. coli strains and contained the toxin genesStx2 or Stx2e, both from phylogroup B1. Our data showed that EPEC was the most frequent pathovar and B1 and E were the most common phylogroups detected in E. coli isolated from the feces of diarrheic and non-diarrheic dogs.

The incidence of Clostridioides difficile and Clostridium perfringens netF-positive strains in diarrheic dogs

The aim of this study was to examine the incidence of Clostridioides (previously Clostridium) difficile and Clostridium perfringens in the feces of diarrheic and non-diarrheic dogs. Also, the presence of other common canine enteropathogens was examined. Toxigenic C. difficile and C. perfringens positive for the NetF-encoding gene (netF) were detected in 11 (11.9%) and seven (7.6%) diarrheic dogs, respectively. Three dogs were diagnosed simultaneously with toxigenic C. difficile and netF-positive C. perfringens. Among other enteropathogens, Giardia sp. was the most common agent detected in dogs positive for toxigenic C. difficile or netF-positive C. perfringens. The results suggest that C. difficile and C. perfringens occur more frequently as a primary cause of diarrhea.

Multilaboratory Survey To Evaluate Salmonella Prevalence in Diarrheic and Nondiarrheic Dogs and Cats in the United States between 2012 and 2014.

ABSTRACTEleven laboratories collaborated to determine the periodic prevalence of Salmonella in a population of dogs and cats in the United States visiting veterinary clinics. Fecal samples (2,965) solicited from 11 geographically dispersed veterinary testing laboratories were collected in 36 states between January 2012 and April 2014 and tested using a harmonized method. The overall study prevalence of Salmonella in cats (3 of 542) was <1%. The prevalence in dogs (60 of 2,422) was 2.5%. Diarrhea was present in only 55% of positive dogs; however, 3.8% of the all diarrheic dogs were positive, compared with 1.8% of the nondiarrheic dogs. Salmonella-positive dogs were significantly more likely to have consumed raw food (P = 0.01), to have consumed probiotics (P = 0.002), or to have been given antibiotics (P = 0.01). Rural dogs were also more likely to be Salmonella positive than urban (P = 0.002) or suburban (P = 0.001) dogs. In the 67 isolates, 27 unique serovars were identified, with three dogs having two serovars present. Antimicrobial susceptibility testing of 66 isolates revealed that only four of the isolates were resistant to one or more antibiotics. Additional characterization of the 66 isolates was done using pulsed-field gel electrophoresis and whole-genome sequencing (WGS). Sequence data compared well to resistance phenotypic data and were submitted to the National Center for Biotechnology Information (NCBI). This study suggests an overall decline in prevalence of Salmonella-positive dogs and cats over the last decades and identifies consumption of raw food as a major risk factor for Salmonella infection. Of note is that almost half of the Salmonella-positive animals were clinically nondiarrheic.

Prevalence and Molecular Characterization of Intestinal Trichomonads in Pet Dogs in East China.

The trichomonad species Tritrichomonas foetus and Pentatrichomonas hominis were recently detected in the feces of dogs with diarrhea. However, little information is available on the prevalence and pathogenicity of these parasites in the canine population. Therefore, the aim of this study was to determine the prevalence and molecular characterization of trichomonads infecting pet dogs in Anhui and Zhejiang provinces, east China. In total, 315 pet dogs, with or without diarrhea, from 7 pet hospitals were included in this epidemiological survey. Microscopy and PCR detected P. hominis in 19.7% (62/315) and 31.4% (99/315) of fecal samples, respectively. T. foetus infection was detected in 0% (0/315) of samples with microscopy and in 0.6% (2/315) with PCR. The prevalence of P. hominis was significantly higher in young dogs (≤12 months) than in adult dogs (>12 months), and was significantly higher in diarrheic dogs (50.6%) than in non-diarrheic dogs (24.3%; P<0.05). Infection with T. foetus did not correlate with any risk factors evaluated in this study. A sequence analysis of the P. hominis PCR products showed minor allelic variations between our sequences and those of P. hominis strains from other hosts in different parts of the world. Type CC1 was the most common strain in dogs in east China. The internal transcribed spacer 1 (ITS1)-5.8S rRNA gene sequences from the 2 T. foetus isolates detected in this study displayed 100% identity and were homologous to the sequences of other strains isolated from domestic cats in other countries.

Detection of Lawsonia intracellularis fecal shedding in dogs in Minas Gerais, Brazil

&lt;p&gt;&lt;bold&gt;Lawsonia intracellularis&lt;/bold&gt;is an obligate intracellular bacterium that is responsible for proliferative enteropathy, an enteric disease endemic in swine and common in foals. However, few studies have investigated this disease in dogs, and there are no reports of dogs infected with&lt;bold&gt; L. intracellularis&lt;/bold&gt;in Latin America. The aim of this study was to evaluate the fecal shedding of&lt;bold&gt; L. intracellularis&lt;/bold&gt;in diarrheic and non-diarrheic dogs in Minas Gerais, Brazil. A total of 58 dogs, 18 apparently healthy and 40 diarrheic, were examined in this study. DNA extracted from feces was analyzed using a nested PCR reaction to detect&lt;bold&gt; L. intracellularis.&lt;/bold&gt;Three out of 40 (7.5%) diarrheic samples, all from 3-month-old puppies, were positive for&lt;bold&gt; L. intracellularis&lt;/bold&gt;. These results highlight the need for additional studies to examine the role of this pathogen as a possible cause of enteric disease in dogs&lt;/p&gt;

Genotypes and antibiotic resistance of canine Campylobacter jejuni isolates

Campylobacter jejuni is the most important cause of bacterial gastroenteritis in humans. It is a commensal in many wild and domestic animals, including dogs. Whereas genotypes of human and chicken C. jejuni isolates have been described in some detail, only little information on canine C. jejuni genotypes is available. To gain more information on genotypes of canine C. jejuni and their zoonotic potential, isolates from routine diagnostics of diarrheic dogs as well as isolates of a prevalence study in non-diarrheic dogs were analyzed. Prevalence of thermophilic Campylobacter among non-diarrheic dogs was 6.3% for C. jejuni, 5.9% for Campylobacter upsaliensis and 0.7% for Campylobacter coli. The C. jejuni isolates were genotyped by multi locus sequence typing (MLST) and flaB typing. Resistance to macrolides and quinolones was genetically determined in parallel. Within the 134 genotyped C. jejuni isolates 57 different sequence types (ST) were found. Five STs were previously unrecognized. The most common STs were ST-48 (11.2%), ST-45 (10.5%) and ST-21 (6.0%). Whereas no macrolide resistance was found, 28 isolates (20.9%) were resistant to quinolones. ST-45 was significantly more prevalent in diarrheic than in non-diarrheic dogs. Within the common time frame of isolation 94% of the canine isolates had a ST that was also found in human clinical isolates. In conclusion, prevalence of C. jejuni in Swiss dogs is low but there is a large genetic overlap between dog and human isolates. Given the close contact between human and dogs, the latter should not be ignored as a potential source of human campylobacteriosis.

Detection of toxins A/B and isolation of Clostridium difficile and Clostridium perfringens from dogs in Minas Gerais, Brazil

The objective of this study was to detect C. difficile A/B toxins and to isolate strains of C. perfringens and C. difficile from diarrheic and non-diarrheic dogs in Brazil. Stool samples were collected from 57 dogs, 35 of which were apparently healthy, and 22 of which were diarrheic. C. difficile A/B toxins were detected by ELISA, and C. perfringens and C. difficile were identified by multiplex PCR. C. difficile A/B toxins were detected in 21 samples (36.8%). Of these, 16 (76.2%) were from diarrheic dogs, and five (23.8%) were from non-diarrheic dogs. Twelve C. difficile strains (21.1%) were isolated, of which ten were A+B+ and two were A−B−. All non-toxigenic strains were isolated from non-diarrheic animals. The binary toxin gene cdtB was found in one strain, which was A+B+ and was derived from a non-diarrheic dog. C. perfringens strains were isolated from 40 samples (70.2%). Of these, 18 (45%) were from the diarrheic group, and 22 (55%) belonged to the non-diarrheic group. All isolates were classified as C. perfringens type A and there was an association between the detection of the cpe gene and the presence of diarrhea. Interestingly, ten strains (25%) were positive for the presence of the cpb2 gene. The high rate of detection of the A/B toxins in non-diarrheic dogs suggests the occurrence of subclinical disease in dogs or carriage of its toxins without disease. More studies are needed to elucidate the epidemiology of C. difficile and C. perfringens in dogs and to better our understanding of C. difficile as a zoonotic agent. This is the first study to report the binary toxin gene in C. difficile strains isolated from dogs in Brazil.

Genotype identification and prevalence of Giardia duodenalis in pet dogs of Guangzhou, Southern China

Giardia duodenalis is a flagellated parasite and is considered one of the most common causes of protozoal diarrhea in both humans and animals worldwide. This paper represents the first study of the prevalence of G. duodenalis in pet dogs in Guangzhou, China. Faecal samples (209 specimens) were obtained from young (<6 months old), adult (6 months to 3 years) and elder dogs (>3 years old). 8.61% (18/209) faecal samples were recorded positive using microscopy examination, and 11.00% (23/209) using PCR. The prevalence was significantly higher in diarrheic dogs (26.31%) compared with non-diarrheic dogs (5.10%), while it was higher in young (25.58%) than both adult (7.37%) and elder (7.04%) dogs and the difference was statistically significant (P<0.05). The prevalence in male dogs 11.30% (13/115) was higher than females 10.87% (10/92), and in suburban dogs (12.15%) higher than urban 9.80%, but the difference was not statistically significant (P>0.05). Sequence analysis of the 23 PCR-positive samples revealed the presence of Assemblage D (18/23), and zoonotic Assemblage A (5/23). The present investigation reported a high infection rate of G. duodenalis in pet dogs, especially in young dogs. Genotypic characterization demonstrated that the zoonotic Assemblage A was found, a fact that poses a potential risk of G. duodenalis transmission from pet dogs to humans. It is suggested that pet owners should take appropriate hygiene measures to prevent and control giardiasis in this region.

Characterization of atypical Enteropathogenic Escherichia coli (aEPEC) isolated from dogs

Enteropathogenic Escherichia coli (EPEC), an important human pathogen has the ability to form attaching and effacing lesions on the intestinal epithelium and has been isolated from a wide range of species. Two EPEC subgroups are recognized: typical (tEPEC) and atypical (aEPEC) strains, differing by the presence of EAF plasmid and bundle-forming pilus (BFP) in typical strains and their absence in atypical strains. This study searched the presence of EPEC strains in 101 fecal samples of diarrheic (n=65) and non-diarrheic (n=36) dogs from two cities in Rio de Janeiro State, Brazil. The isolates were evaluated for the presence of eae, tir, espA, espB and bfpA genes, EAF plasmid, and for the insertion site of the LEE locus. Cell-adherence assays, fluorescent actin staining (FAS) test, hemolysin production and serotyping were also performed. Twenty eight aEPEC isolates were recovered from 48 eae-positive fecal samples, 24 from diarrheic animals and 4 from non-diarrheic ones. PCR showed that most isolates was positive for β or γ intimin, and for β or α subtypes of tir, espA and espB. Six isolates showed a selC insertion of locus LEE. Only two isolates from the same diarrheic animal harbored the bfpA gene, and none presented the EAF plasmid. Most isolates was FAS-positive and showed a localized adherence-like (LAL) in a 6h HeLa cell-adherence assay. A wide diversity of serotypes was detected including O4:H16 and O51:H40, previously described in human disease. The phenotypic and genotypic markers of aEPEC isolated from diarrheic and non-diarrheic dogs were similar to those found in isolates recovered from human disease.

Evaluation of Five Enzyme Immunoassays Compared with the Cytotoxicity Assay for Diagnosis of Clostridium Difficile-Associated Diarrhea in Dogs

Clostridium difficile-associated-diarrhea (CDAD) is a nosocomial infection in dogs. Diagnosis of this infection is dependent on clinical signs of disease supported by laboratory detection of C. difficile toxins A or B, or both, in fecal specimens via enzyme-linked immunosorbent assay (ELISA). Unfortunately, to the authors' knowledge, commercially available ELISAs have not been validated in dogs to date. We evaluated 5 ELISAs done on 143 canine fecal specimens (100 diarrheic and 43 nondiarrheic dogs) and on 29 C. difficile isolates. The results of each ELISA were compared with the cytotoxin B tissue culture assay (CTA). Clostridium difficile was isolated from 23% of the fecal specimens. Eighteen of the 143 fecal specimens were toxin positive (15 diarrheic and 3 nondiarrheic dogs). On the basis of multiplex polymerase chain reaction (PCR) analysis for toxin-A and -B genes, 72% of the isolates were toxigenic. The carriage rate of toxigenic isolates in diarrheic dogs was higher than that in the nondiarrheic dogs; however, these differences were not statistically significant. A good correlation was found between CTA, PCR, and culture results. The ELISAs done on fecal specimens collected from diarrheic dogs had low sensitivity (7-33%). In contrast, ELISA for toxin A or B, or both, performed on toxigenic isolates had high sensitivity (93%). These results suggest that commercially available human ELISAs are inadequate for the diagnosis of canine C. difficile-associated diarrhea when tested on fecal specimens. In contrast, the Premier ToxinA/B and Techlab ToxinA/B ELISAs may be useful for the diagnosis of canine CDAD when used on toxigenic isolates.