Diarrheic Fecal Samples Research Articles

Prevalence and molecular characterisation of Giardia duodenalis in calves with diarrhoea

The aim of this study was to investigate the prevalence of Giardia duodenalis in diarrhoeal faeces from calves that were reared in Korea using PCR and ELISA. Diarrhoeal faecal samples...

Genetic heterogeneity of the VP6 gene and predominance of G6P[5] genotypes of Brazilian porcine rotavirus C field strains

The aim of this study was to investigate the genetic diversity of the VP6, VP7, and VP4 genes of 15 Brazilian wild-type porcine RVC strains identified in diarrheic fecal samples. The VP6 gene analysis demonstrated heterogeneity between the 15 RVC strains, which clustered in three distinct genotypes (I1, I5, and I6). In the VP7 and VP4 gene analysis, the genotype combination G6P[4] was detected in only one strain (UEL-77), while G6P[5] was the most commonly (n = 14) detected in RVC strains identified in the Brazilian pig herds evaluated, indicating its probable predominance in this country, mainly in 2014.

The role of human adenoviruses type 41 in acute diarrheal disease in Minas Gerais after rotavirus vaccination.

Human adenovirus species F (HAdV-F) type 40 and 41 are commonly associated with acute diarrheal disease (ADD) across the world. Despite being the largest state in southeastern Brazil and having the second largest number of inhabitants, there is no information in the State of Minas Gerais regarding the role of HAdV-F in the etiology of ADD. This study was performed to determine the prevalence, to verify the epidemiological aspects of infection, and to characterize the strains of human adenoviruses (HAdV) detected. A total of 377 diarrheal fecal samples were obtained between January 2007 and August 2011 from inpatient and outpatient children of age ranging from 0 to 12 years. All samples were previously tested for rotavirus, norovirus, and astrovirus, and 314 of 377 were negative. The viral DNA was extracted, amplified using the polymerase chain reaction and the HAdV-positive samples were sequenced and phylogenetically analyzed. Statistical analyses were performed using the Chi-square test (p < 0.05), considering two conditions: the total of samples tested (377) and the total of negative samples for the remaining viruses tested (314). The overall prevalence of HAdV was 12.47% (47/377); and in 76.60% (36/47) of the positive samples, this virus was the only infectious agent detected. The phylogenetic analysis of partial sequences of 32 positive samples revealed that they all clustered with the HAdV-F type 41. The statistical analysis showed that there was no correlation between the onset of the HAdV infection and the origin of the samples (inpatients or outpatients) in the two conditions tested: the total of samples tested (p = 0.598) and the total of negative samples for the remaining viruses tested (p = 0.614). There was a significant association in the occurrence of infection in children aged 0–12 months for the condition 1 (p = 0.030) as well as condition 2 (p = 0.019). The occurrence of infections due to HAdV did not coincide with a pattern of seasonal distribution. These data indicate the significant involvement of HAdV-F type 41 in the etiology of ADD in Minas Gerais, which demonstrates the importance of other viral agents in the development of the disease after the introduction of rotavirus vaccine immunization.

Molecular detection of bovine coronavirus in a diarrhea outbreak in pasture-feeding Nellore steers in southern Brazil.

Worldwide diarrhea outbreaks in cattle herds are more frequently detected in calves being that diarrhea outbreaks in adult cattle are not common. Winter dysentery (WD) is a bovine coronavirus (BCoV) enteric infection that is more reported in Northern hemisphere. Seasonal outbreaks of WD in adult cattle occur mainly in dairy cows. WD has not been described in beef cattle herds of tropical countries. This study describes the molecular detection of BCoV in a diarrhea outbreak in beef cattle steers (Nellore) raised on pasture in Parana, southern Brazil. During the outbreak, the farm had about 600 fattening steers. Watery and bloody diarrhea unresponsive to systemic broad-spectrum antibiotic therapy reveals a morbidity rate of approximately 15 %. The BCoV N gene was identified in 42.9 % (6/14) of the diarrheic fecal samples evaluated by semi-nested polymerase chain reaction (SN-PCR) technique. Other enteric microorganisms occasionally identified in adult cattle and evaluated in this study such as bovine groups A, B, and C rotavirus, bovine viral diarrhea virus, bovine torovirus, aichivirus B, and Eimeria sp. were not identified in the fecal samples. To the best knowledge of the authors, this is the first description of the BCoV diagnosis in fecal samples collected in a diarrhea outbreak in adult beef cattle grazing in the grass in a tropical country.

Viral metagenomics analysis of picobirnavirus-positive feces from children with sporadic diarrhea in China.

Picobirnaviruses (PBVs) infect humans and a wide range of animals and may cause diarrhea. The aim of this study was to investigate PBV infection and its association with diarrhea. Here, seven PBV RT-PCR-positive fecal samples from diarrheic children and four fecal samples from healthy children as controls were analyzed by viral metagenomics. The results indicated that all the seven diarrheic fecal samples contain high titers of PBV sequences, while three of the controls were negative, and one had low titers of PBV. Three of the diarrheic fecal samples were also positive for other viruses, including anellovirus, human gyrovirus, human parechovirus, and porcine stool-associated circular virus. PBV sequences from the seven patients were assembled, generating seven large contigs with the complete ORF of RNA-dependent RNA polymerase (RdRp). Phylogenetic analysis of the amino acid sequences of RdRp indicated that the seven PBVs in the present study belonged to three different genogroups. Our data suggest that PBV might have been the cause of diarrhea in these seven children.

Antimicrobial assay of shigatoxigenic E. coli (STEC) and enteropathogenic E. coli (EPEC) isolated from diarrhoeic faecal samples of piglets and infants in Mizoram

Antimicrobial resistance is a common problem which is accelerating day by day in veterinary medicine. The main reason is believed to be the indiscriminate and irrational use of antibiotics. Diarrhoeic faecal samples (584: 320 from piglets and 264 from infants) were collected from different farms and hospitals located in different districts of Mizoram. Out of 1,260 E. coli isolates, 65 (5.15%) E. coli isolates were found positive for at least 1 virulence gene (stx1, stx2, eaeA, hlyA) under the study, of which 40 (3.17%) and 25 (1.98%) were recorded as STEC and EPEC, respectively, when screened by multiplex PCR. All the 65 E. coli isolates were subjected to antimicrobial sensitivity test against 12 commonly used antimicrobial agents. Among the isolates from piglets, highest sensitivity was exhibited by chloramphenicol (89.58%) and highest resistance by nalidixic acid (85.41%). On the other hand, enrofloxacin exhibited 100% sensitivity while amoxicillin, polymyxin B and kanamycin exhibited 100% resistance among the infant isolates. It may be concluded that 5.15% isolates were positive for virulence and the isolates showed increased tendency of resistance to many of the commonly used antibiotics reflecting a greater threat to treat the commonly occurring diseases with antibiotics.

Hospital-based Clostridium difficile infection surveillance reveals high proportions of PCR ribotypes 027 and 176 in different areas of Poland, 2011 to 2013.

As part of the European Clostridium difficile infections (CDI) surveillance Network (ECDIS-Net), which aims to build capacity for CDI surveillance in Europe, we constructed a new network of hospital-based laboratories in Poland. We performed a survey in 13 randomly selected hospital-laboratories in different sites of the country to determine their annual CDI incidence rates from 2011 to 2013. Information on C. difficile laboratory diagnostic testing and indications for testing was also collected. Moreover, for 2012 and 2013 respectively, participating hospital-laboratories sent all consecutive isolates from CDI patients between February and March to the Anaerobe Laboratory in Warsaw for further molecular characterisation, including the detection of toxin-encoding genes and polymerase chain reaction (PCR)-ribotyping. Within the network, the mean annual hospital CDI incidence rates were 6.1, 8.6 and 9.6 CDI per 10,000 patient-days in 2011, 2012, and 2013 respectively. Six of the 13 laboratories tested specimens only on the request of a physician, five tested samples of antibiotic-associated diarrhoea or samples from patients who developed diarrhoea more than two days after admission (nosocomial diarrhoea), while two tested all submitted diarrhoeal faecal samples. Most laboratories (9/13) used tests to detect glutamate dehydrogenase and toxin A/B either separately or in combination. In the two periods of molecular surveillance, a total of 166 strains were characterised. Of these, 159 were toxigenic and the majority belonged to two PCR-ribotypes: 027 (n=99; 62%) and the closely related ribotype 176 (n=22; 14%). The annual frequency of PCR-ribotype 027 was not significantly different during the surveillance periods (62.9% in 2012; 61.8% in 2013). Our results indicate that CDIs caused by PCR-ribotype 027 predominate in Polish hospitals participating in the surveillance, with the closely related 176 ribotype being the second most common agent of infection.

Prevalence of Balantidium coli infection in sows in Hunan province, subtropical China.

Balantidium coli is a cosmopolitan parasitic-opportunistic pathogen that can be found throughout the world. However, little information is available about prevalence of B. coli in pigs in China. In the present study, the prevalence of B. coli in pigs was investigated in Hunan province, subtropical China, between January 2012 and August 2014. A total of 3925 diarrheic fecal samples from nine representative administrative regions in Hunan province, subtropical China, were examined for the presence of B. coli cysts and/or trophozoites using microscopy after sedimentation with water. The overall prevalence of B. coli in pigs was 36.9% (1450/3925). The present survey revealed high circulation of B. coli in pigs in Hunan province, subtropical China, which poses potential threats to human health. The results of the present investigation have important implications for the control of B. coli infections in pigs in Hunan province, subtropical China. To our knowledge, this is the first comprehensive report of B. coli prevalence in sows in Hunan province, subtropical China.

Concurrent Infection of Rotavirus and Shiga ToxigenicEscherichia coli(STEC) in Buffalo Calves in Mumbai Metropolis, Western India

Faecal samples from 113 diarrhoeic cattle and buffalo calves less than one year of age, from different farms in Mumbai metropolis in India, were screened for the presence of Rotavirus and Shiga toxin-producing Escherichia coli (STEC). RNA-Poly acrylamide gel electrophoresis (RNA-PAGE) analysis of these samples revealed 11 (9.73%) as positive for group A rotavirus (GARV). Out of 11 samples found positive by RNA-PAGE, which were subjected to amplification of VP7 gene of GARV, 10 samples yielded a specific product of 1, 013 bp. The incidence of Shiga toxigenic E. coliin diarrhoeic buffalo calves was 13.23% (9 out of 68) and 6.66% in the cattle calves (3 out of 45) as demonstrated by multiplex polymerase chain reaction (PCR) of the shiga toxin (stx) genes. Eight (66.66%) isolates carried stx2 gene and four (33.33%) isolates had stx1 gene. To the best of our knowledge, we report for the first time the concurrent infection of Shiga toxigenic E. coli and GARV (6 cases; 8.82%) in diarrhoeic faecal samples of buffalo calves from India.

Phylogenetic analysis of a G6P[5] bovine rotavirus strain isolated in a neonatal diarrhea outbreak in a beef cattle herd vaccinated with G6P[1] and G10P[11] genotypes.

The aim of this study was to perform the molecular characterization of the eleven genes of a G6P[5] bovine group A rotavirus (RVA) strain detected in a diarrhea outbreak from a vaccinated beef cattle herd. The outbreak affected 80 % of calves between 15–30 days old. RVA was identified by RT-PCR in 12 (70.6 %) out of 17 diarrheic fecal samples evaluated. The rotavirus wild-type strain had the genotype constellation G6(IV)-P[5](IX)-I2c-R2-C2-M2-A3-N2-T6-E2e-H3a. This study confirms the importance of homotypic immunity against the bovine RVA P[5] genotype in neonatal diarrhea in cattle herds that are regularly vaccinated against rotaviruses.

Detection of Torque teno sus virus in diarrheic piglet fecal samples positive or negative for porcine group A rotavirus

Association of Torque teno sus virus (TTSuV) and porcine group A rotavirus (PoRVA) was evaluated in PoRVA-positive or PoRVA-negative diarrheic piglet fecal samples. Molecular TTSuV detection was 40.4% (21/52) and 53.3% (49/92) in PoRVA-positive and -negative fecal samples, respectively. No association (P = .19) was observed between TTSuV and PoRVA diarrhea.

Detection of Campylobacter in human faecal samples in Fiji.

Data on campylobacteriosis in developed countries are well documented; in contrast, few studies on campylobacteriosis have been conducted in developing countries. This study was undertaken to test for Campylobacter in human faecal samples sent to the two major pathology laboratories in Fiji. A total of 408 diarrhoeal faecal samples were collected from the two major hospital pathology laboratories in Central Fiji (Suva) and Western Fiji (Lautoka) between December 2012 and February 2013 and from June to July 2013. Samples were analysed for the presence of Campylobacter using polymerase chain reaction (PCR) based methods. Campylobacter was detected in 241/408 (59.1%) of samples tested using PCR. Samples from children aged less than five accounted for 21.6% of positive cases. Campylobacter was detected in 59.1% of diarrhoeal samples collected from the two main laboratories in Fiji. A high proportion of children under five years with Campylobacter has been reported in other countries and could be due to parents being more likely to seek medical attention. Further studies are required to confirm the species of Campylobacter that are predominantly associated with gastroenteritis in Fiji.

VP6 gene diversity in 11 Brazilian strains of porcine group C rotavirus.

Porcine group C rotavirus (RVC) is recognised as an enteric pathogen in piglets worldwide. The VP6 gene of RVC is divided into seven I-genotypes. Genotypes I2 and I3 are found in human and bovine strains, respectively; the porcine strains are divided into the other five genotypes (I1, I4-I7). In this study, molecular analysis of nearly the full length of the VP6 gene was performed in 11 Brazilian wild-type porcine RVC strains identified in diarrhoeic faecal samples, which were collected from eight pig farms located in five Brazilian states from piglets of 1-4 weeks of age. The nucleotide sequences of the VP6 gene showed 82.9-100 % identity between the Brazilian strains, 84.9-93.1 % with the prototype Cowden strain, and 82.4-92.2 % with other porcine RVC strains. In the 11 diarrhoeic faecal samples analysed in this study, three distinct porcine RVC genotypes (I1, I5, and I6) were identified and none were predominant. The results presented in this study revealed a high nucleotide diversity of the VP6 gene in porcine RVC field strains circulating in Brazil, which highlights the importance of further epidemiological and molecular surveys worldwide.

Prevalence of Cryptosporidium-associated diarrhea in a high altitude-community of Saudi Arabia detected by conventional and molecular methods.

Cryptosporidium diarrhea represents a relevant clinical problem in developing countries. In Al-Taif, a city of Saudi Arabia that lies at an altitude of an around 2 km above the sea level, Cryptosporidium infection seems to be undiagnosed in nearly all clinical laboratories. Furthermore, nothing was published regarding Cryptosporidium-associated diarrhea in this area. The objectives of this research were to (1) determine the Cryptosporidium prevalence among patients with diarrhea and (2) to estimate the performances of 3 different diagnostic methods. Total 180 diarrheal fecal samples, 1 sample per patient, were collected between January and August 2013. Samples were screened for Cryptosporidium with modified Zeihl Neelsen (ZN) microscopy, RIDA® Quick lateral flow (LF) immunotest, and a previously published PCR. The Cryptosporidium prevalence rate was 9.4% (17/180), 10% (18/180), and 11.6% (21/180) by microscopy, LF, and PCR test, respectively. Infection was significantly (P=0.004) predominant among children <5 years (22%) followed by children 5-9 years (11.1%). Although infection was higher in males than in females (16.2% males and 8.5% females), the difference was not statistically significant (P=0.11). Compared to PCR, the sensitivity of microscopy and the LF test were 80.9%, 85.7%, respectively. To conclude, high Cryptosporidium-associated diarrhea was found in this area especially in children ≤9 years. The PCR test showed the best performance followed by the LF test and ZN staining microscopy. The primary health care providers in Al-Taif need to be aware of and do testing for this protozoon, particularly for children seen with diarrhea.

Prevalence and identification of Cryptosporidium species in paediatric patients with diarrhoea.

To determine the prevalence of Cryptosporidium infection in children with diarrhoea, identify associated factors and identify the parasite using Polymerase Chain Reaction (PCR). A total of 138 diarrhoeic faecal samples were collected between August 2011 and February 2013, from children under 12 years of age, admitted to paediatric units of Teaching Hospitals, Kandy and Peradeniya, Sirimawo Bandaranayake Childrens' Hospital, Peradeniya and District General Hospital, Matale. One hundred faecal samples collected from healthy children were used as controls. All control and test samples were screened for the presence of Cryptosporidium oocysts with Modified Ziehl-Neelsen (MZN) method and PCR. Prevalence of Cryptosporidium infection among children with diarrhoea was 5.7%. All the cases positive for Cryptosporidium were below 3 years of age. The majority (7 out of 8) of the positive cases had watery diarrhoea while none of the healthy children excreted Cryptosporidium oocysts in the faeces. Of the 8 positive cases, 6 had a history of animal contact. A large proportion of positive cases used pipe borne municipal water. The majority (66.6%) of positive cases did not consume boiled cooled water. We were able to identify C. parvum from one of the eight cases that had diarrhoea. The current study shows that Cryptosporidium is one of the aetiological agents responsible for childhood watery diarrhoea in Sri Lanka, thus stressing the importance of routine stool examination for Cryptosporidium oocysts. This study recommends boiling water as an important measure to prevent the transmission of Cryptosporidium oocysts. Further molecular studies are needed to determine the other species of Cryptosporidium responsible for cryptosporidiosis in children in Sri Lanka.

Severe diarrhea outbreak in beef calves (Bos indicus) caused by G6P[11], an emergent genotype of bovine rotavirus group A

The episodes of diarrhea caused by neonatal bovine rotavirus group A (BoRVA) constitute one of the major health problems in the calf rearing worldwide. The main G (VP7) and P (VP4) genotypes of BoRVA strains involved in the etiology of diarrhea in calves are G6P[1], G10P[11], G6P[5], and G8P[1]. However, less frequently, other G and P genotypes have been described in BoRVA strains identified in diarrheic fecal samples of calves. This study describes the identification and molecular characterization of an emerging genotype (G6P[11]) in BoRVA strains involved in the etiology of a diarrhea outbreak in beef calves in a cattle herd of high production in extensive management system. The diarrhea outbreak, which showed high morbidity (60%) and lethality (7%) rates, occurred in calves (n= 384) Nelore (Bos indicus) up to 30-day-old from the State of Mato Grosso do Sul, Brazil. BoRVA was identified in 80% (16/20) of the fecal samples analyzed by polyacrylamide gel electrophoresis (PAGE) technique. In all PAGE-positive fecal samples were amplified products with 1,062-bp and 876-bp in the RT-PCR assays for VP7 (G type) and VP4 (VP8*) (P type) of BoRVA, respectively. The nucleotide sequence analysis of VP7 and VP4 genes of four wild-type BoRVA strains showed G6-III P[11]-III genotype/lineage. The G6P[11] genotype has been described in RVA strains of human and animal hosts, however, in calves this genotype was only identified in some cross-sectional studies and not as a single cause of diarrhea outbreaks in calves with high morbidity and lethality rates as described in this study. The monitoring of the G and P genotypes of BoRVA strains involved in diarrhea outbreaks in calves is important for both animal and public health by allowing the identification of the most frequent genotypes, the characterization of novel genotypes and to identify reassortments with genotypes described in animal and human hosts. The results of this study show the importance of the monitoring of the genotypes of BoRVA strains involved in episodes of bovine neonatal diarrhea as for characterization of frequency of occurrence and pathogenic potential of uncommon genotypes as for monitoring of the emergency of different BoRVA genotypes not included in commercial vaccines.

Characterization of Escherichia coli virulence genes, pathotypes and antibiotic resistance properties in diarrheic calves in Iran.

BackgroundCalf diarrhea is a major economic concern in bovine industry all around the world. This study was carried out in order to investigate distribution of virulence genes, pathotypes, serogroups and antibiotic resistance properties of Escherichia coli isolated from diarrheic calves.ResultsTotally, 76.45% of 824 diarrheic fecal samples collected from Isfahan, Chaharmahal, Fars and Khuzestan provinces, Iran were positive for E. coli and all of them were also positive for cnf2, hlyA, cdtIII, f17c, lt, st, stx1, eae, ehly, stx2 and cnf1 virulence genes. Chaharmahal had the highest prevalence of STEC (84.61%), while Isfahan had the lowest (71.95%). E. coli serogroups had the highest frequency in 1–7 days old calves and winter season. Distribution of ETEC, EHEC, AEEC and NTEC pathotypes among E. coli isolates were 28.41%, 5.07%, 29.52% and 3.49%, respectively. Statistical analyses were significant for presence of bacteria between various seasons and ages. All isolates had the high resistance to penicillin (100%), streptomycin (98.25%) and tetracycline (98.09%) antibiotics. The most commonly detected resistance genes were aadA1, sul1, aac[3]-IV, CITM, and dfrA1. The most prevalent serogroup among STEC was O26.ConclusionsOur findings should raise awareness about antibiotic resistance in diarrheic calves in Iran. Clinicians should exercise caution when prescribing antibiotics.

Isolation and identification of diarrheagenic &lt;i&gt;Escherichia coli&lt;/i&gt; causing colibacillosis in calf in selective areas of Bangladesh

This study was carried out to isolate, identify and characterize Escherichia coli from diarrheic calves, their prevalence and antibiotic sensitivity pattern during the period from November 2011 to December 2012. For this purpose 100 diarrheic fecal samples were collected from four different areas of Bangladesh. The samples were aseptically collected and brought to the laboratory of Medicine, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh. The organisms were identified as E. coli on the basis of cultural, morphological and biochemical characteristics. Further the isolated bacteria were confirmed by amplification of E. coli specific 16sRNA gene. The antibiotic resistant pattern of the isolates was also performed. Out of 100 samples 49 (49%) were found to be positive for E. coli. The highest prevalence was found in Shahjadpur, Sirajganj (53.33%) while lowest was in BAU, Mymensingh (40%). The antibiogram study revealed that all isolates were resistant against ampicillin, erythromycin, azithromycin and streptomycin where erythromycin was 100% resistant, whereas 100% sensitivity was found to tetracycline and gentamicin which might be the best choice of drug for the treatment of diarrheagenic E. coli causing colibacillosis in calves.DOI: http://dx.doi.org/10.3329/bjvm.v11i2.19139Bangl. J. Vet. Med. (2013).11(2): 145-149

Molecular epidemiology of Porcine torovirus (PToV) in Sichuan Province, China: 2011-2013.

BackgroundPorcine torovirus (PToV) is a member of the genus Torovirus which is responsible for gastrointestinal disease in both human beings and animals with particular prevalence in youth. Torovirus infections are generally asymptomatic, however, their presence may worsen disease consequences in concurrent infections with other enteric pathogens.MethodsA total of 872 diarrheic fecal samples from pigs of different ages were collected from 12 districts of Sichuan Province in the southwest of China. RT-PCR was done with PToV S gene specific primers to detect the presence of PToV positive samples. M gene specific primers were used with the PToV positive samples and the genes were sequenced. A phylogenetic tree was constructed based on the M gene nucleotide sequences from the 19 selected novel Sichuan strains and 21 PToV and BToV M gene sequences from GenBank.ResultsA total of 331 (37.96%, 331/872) samples were found to be positive for PToV and the highest prevalence was observed in piglets aged from 1 to 3 weeks old. Through phylogenetic inference the 40 PToV M gene containing sequences were placed into two genotypes (I & II). The 19 novel Sichuan strains of genotype I showed strong correlations to two Korean gene sequences (GU-07-56-11 and GU-07-56-22). Amino-acid sequence analysis of the 40 PToV M gene strains revealed that the M gene protein was highly conserved.ConclusionsThis study uncovered the presence of PToV in Sichuan Province, and demonstrated the need for continuous surveillance PToV of epidemiology.

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.