Fecal ammonia, urea, volatile fatty acid and lactate levels in dairy cows and their pathophysiological significance during diarrhea

Normal fecal samples were taken from lactating cows fed either a total mixed ration (TMR; n = 30) or pasture‐based diet (20) and from dry cows fed mainly on hay (15). Diarrheic fecal samples (n = 51) were collected from 21 sick dairy cows. Fecal analyses of ammonia, urea, lactate and volatile fatty acid (VFA) levels were used to evaluate colonic fermentation. Most normal feces had reasonably neutral pH, however, alkaline feces were observed in diarrheic cows. Although fecal lactate is higher in cows on grazing pasture, lactate levels were generally lower in the cows in the present study. Fecal VFA levels were higher in lactating cows than in dry cows. Elevated fecal urea was observed in diarrheic cows, however, many fecal samples in normal and diarrheic cows contained no urea. Fecal VFA levels in diarrheic cows were lower than in normal lactating cows, but were approximately equivalent to those in dry cows. Grazing or dry cows showed higher acetate and lower n‐butyrate proportions compared with TMR‐fed or diarrheic cows. Higher proportions of branched chain VFAs were observed in diarrheic cows, and the lowest level was observed in grazing cows. The present results indicate that intracolonic nitrogen equilibrium and proteolytic fermentation are altered by diarrheic status.     

Fecal D- and L- lactate, succinate, and volatile fatty acid levels in young dairy calves

For evaluation of physiologically significant organic anions in the colonic environment, 87 samples of normal feces were collected from the rectum of 15 calves less than 60 days old. The calves were fed milk replacer with free access to starter diet and hay. After fecal extraction with water, pH, D- and L-lactate, succinate, and volatile fatty acid (VFA) concentrations were determined. There was wide variation in fecal pH (4.3 to 7.7). Higher lactate concentrations were observed in feces samples with lower pH, and most of these samples were collected during the first 4 weeks of life. Elevated lactate concentrations included both the D- and L-isomers, and the D-isomer comprised approximately 30-50% of total lactate. Elevated succinate concentrations were observed in only 8 fecal samples, while other samples had lower or trace amounts of succinate. Elevated fecal succinate showed no relationship with fecal pH or VFA concentrations. Fecal VFA concentrations were lower in samples collected in the early postnatal stage, but fecal VFA concentrations were not necessarily related to age. We confirmed that fecal D- and L-lactate concentrations increased with a concomitant decrease of VFA in the acidic lumen of the colon, although acidic feces were not necessarily accompanied by elevated concentrations of lactate. In contrast, succinate production was not related to fecal pH or VFA concentrations.     

Fecal excretion of alcohols and organic anions in neonatal dairy calves

To clarify colonic fermentation during the perinatal period, 22 dairy calves less than 6 weeks old were used. They were given a milk replacer following colostrum feeding. A total 100 samples of normal feces including meconium were collected from the rectum of the calves. Fecal pH, alcohols, lactate and volatile fatty acids (VFAs) were analyzed. Higher ethanol and n -propanol concentrations were found in many fecal samples particularly in the first 2 weeks after birth, but these metabolites showed consistently lower concentrations thereafter. By contrast, higher concentrations of methanol were observed in some samples for all ages examined. Fecal VFA increased abruptly within a few days of birth, and mainly consisted of acetate and n -butyrate. During the first 2 weeks, the proportion of n -butyrate in VFAs decreased and that of propionate increased gradually. Proportions of VFAs were almost stable at 3–6 weeks of age (acetate, propionate and n -butyrate in increasing order). Higher concentrations of lactate and lower pHs were observed in the fecal samples during the first 2 weeks, and concentrations decreased thereafter. Accelerated colonic production of ethanol and n -propanol was confirmed during the early 2 weeks, in addition to organic acid fermentation as reported previously.     

Effects of supplementary probiotics to two different diets on dry matter intake, daily gain, digestibility, ruminal pH, and fecal microbial populations and metabolites in ewes

To study the effects of supplementary probiotics on dry matter intake (DMI), daily gain (DG), digestibility, ruminal pH, and fecal microbial populations and metabolites in ruminants, two reversal trials were conducted by using four Suffolk ewes fitted with rumen cannula. The ewes were fed with oat hay and with concentrate and oat hay in the ratio of 60 : 40 in experiments 1 and 2, respectively. The ewes in the treatment groups were supplemented with 10 g/day/head probiotics for 49 days. Fresh fecal samples were collected to measure microbial populations and metabolites. On days 43–47 total feces was collected to measure digestibility, and on the days 48 and 49 ruminal pH was measured. No significant difference of DMI, DG, dry matter digestibility, and ruminal pH was observed between the control and treatment groups. The probiotics treatment tended to increase crude fiber (P = 0.11) and organic cell wall digestibility (P = 0.18). In the final week, probiotics treatment significantly increased the fecal population of Bacilli (P < 0.05) and mold (P < 0.01) in experiment 1 and 2, respectively. No significant difference of fecal VFA and ammonia concentrations between the control and treatment groups was observed. The supplementary probiotics changed population of some microbial strains in the feces and possibly the large intestine of ewes.     

The effect of diet on fecal moisture, osmolarity of fecal extracts, products of bacterial fermentation and loss of minerals in feces of weaned pigs

Litters of pigs were allotted to one of three dietary treatments. Treatment 1 (T1) consisted of a corn-soybean meal starter diet. Pigs fed treatment 2 (T2) received a steamed, rolled oat groats-casein diet and pigs in treatment 3 (T3) remained with the sow. Four pigs/treatment were used to investigate the difference in performance and the cause of post-weaning diarrhea associated with early weaning of pigs at 4 wk of age to a starter diet. Fecal moisture, osmolarity, acetic acid, lactic acid and glucose contents were all good indicators of dietary differences because of treatment X age interactions. These variables increased faster in fecal extracts from pigs fed the corn-soybean meal diet. Lactic acid, volatile fatty acid (VFA) levels, glucose and pH values were indicative of a more active bacterial fermentation in pigs receiving T1 than in those receiving T2 or pigs remaining with the sow (T3). Excess minerals appear to contribute significantly to the osmolarity of fecal material. Of the anions, lactate was the main contributor to the osmolarity of feces of T1 pigs, followed by P, VFA, of which acetic acid contributed 70%, and Cl. The main cations were K, Na and Ca. In T2, P was the main anion, followed by lactate, VFA and Cl, while the main cations were Na, K and Ca. Minerals seemed to be the major osmotic particles in fecal extracts of pigs remaining with the sow. Phosphorus was the major anionic contributor to osmolarity, followed by VFA, Cl and lactic acid. Potassium was the major cation, followed by Na and Ca.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of Maillard reaction products on ruminal and fecal acid-resistant E. coli, total coliforms, VFA profiles, and pH in steers

Six ruminally cannulated Angus-cross steers (362 kg) were used in a replicated 3 x 3 Latin square design to determine effects of supplementing Maillard reaction products (MRP) on acid-resistant E. coli and coliform populations. Steers were fed roughage-based diets supplemented (DM basis) with either 10% soybean meal (SBM), 10% nonenzymatically browned SBM (NESBM), or 10% SBM top-dressed with 45 g of a lysine-dextrose Maillard reaction product (LD-MRP). Equal weights of dextrose, lysine hydrochloride, and deionized water were refluxed to produce the LD-MRP. The NESBM was manufactured by treating SBM with invertase enzyme, followed by heating to induce nonenzymatic browning. Steers were allowed slightly less than ad libitum access to diets fed twice daily and were adapted to their respective treatments within 10 d. On d 11, ruminal and fecal samples were collected at 0, 2, 4, 6, 8, and 12 h after feeding from each of the steers and transported to the laboratory for microbial analysis. Ruminal samples and feces were analyzed for pH and VFA, and both ruminal fluid and feces were tested for acid-resistant E. coli and total coliforms by incubating samples in tryptic soy broth adjusted to pH 2, 4, and 7. Ruminal pH and total VFA concentrations did not differ among treatments. The molar proportion of ruminal acetate was higher (P < 0.05) for steers receiving NESBM than for steers receiving SBM and LD-MRP. At pH 4, steers that received NESBM had lower (P < 0.05) ruminal populations of E. coli and total coliforms than steers that received SBM. No differences were observed for ruminal E. coli and total coliforms at pH 2 and 7. Fecal pH was lower (P < 0.05) for steers fed NESBM than for steers fed SBM or LD-MRP. Molar proportions of fecal acetate were lower (P < 0.05) and proportions of butyrate and isovalerate were higher (P < 0.05) for steers fed NESBM compared with steers fed SBM. Fecal E. coli at pH 4 was lower (P < 0.05) for steers fed NESBM than for steers fed LD-MRP. Fecal E. coli and total coliforms at pH 2 and 7 did not differ among treatments. Dietary MRP had limited effectiveness at decreasing acid-resistant coliforms in the rumen and feces of cattle. Acid resistance in coliforms may depend on protein availability.     

Epidemiologic survey of diarrhea in foals

Epidemiologic and etiologic data about diarrhea in foals were collected under a planned prospective recording and monitoring study. The survey and monitoring procedures included a survey to obtain an overview of current horse management practices on participating farms, a daily health record survey to obtain information on mares and their foals, and collection of feces from 19 of 144 diarrheic foals and 10 age-matched nondiarrheic foals for electron microscopy, ELISA for rotavirus, and bacteriologic culture. Coronavirus was detected in the feces of diarrheic as well as clinically normal foals. Rotavirus was detected in the feces of diarrheic foals only. With regard to agents found in the feces, there was no significant (P less than 0.05) difference between diarrheic and nondiarrheic foals. Half of the 297 foals on which data were available developed diarrhea. Most foals that developed diarrhea lacked other clinical manifestations of disease. Basic cleanliness at foaling was associated with a lower percentage of foals developing diarrhea. Prophylactic use of antibiotics and vitamins in newborn foals was associated with a higher percentage of foals developing diarrhea. A higher percentage of foals born to visiting mares developed diarrhea, compared with foals born to resident mares.     

Effect of trehalose supplementation in milk replacer on the incidence of diarrhea and fecal microbiota in preweaned calves

Trehalose, a nonreducing disaccharide consisting of d-glucose with α,α-1,1 linkage, was evaluated as a functional material to improve the gut environment in preweaned calves. In experiment 1, 173 calves were divided into two groups; the trehalose group was fed trehalose at 30 g/animal/d with milk replacer during the suckling period, and the control group was fed nonsupplemented milk replacer. Medication frequency was lower in the trehalose group (P < 0.05). In experiment 2, calves (n = 20) were divided into two groups (control group [n = 10] and trehalose group [n = 10]) based on their body weight and reared under the same feeding regimens as in experiment 1. Fresh feces were collected from individual animals at the beginning of the trial (average age 11 d), 3 wk after trehalose feeding (experimental day 22), and 1 d before weaning, and the fecal score was recorded daily. Fecal samples were analyzed for fermentation parameters and microbiota. The fecal score was significantly lower in the trehalose group than in the control group in the early stage (at an age of 14 to 18 d; P < 0.05) of the suckling period. Calves fed trehalose tended to have a higher proportion of fecal butyrate on day 22 than calves in the control group (P = 0.08). Population sizes of Clostridium spp. were significantly lower (P = 0.036), whereas those of Dialister spp. and Eubacterium spp. tended to be higher in the feces of calves in the trehalose group on day 22 (P = 0.060 and P = 0.083). These observations indicate that trehalose feeding modulated the gut environment and partially contributed to the reduction in medication frequency observed in experiment 1.     

Bovine salmonellosis: experimental production and characterization of the disease in calves, using oral challenge with Salmonella typhimurium.

A highly virulent strain of Salmonella tyhimurium was given orally to produce disease experimentally in 21 normal colostrum-fed calves 3 to 9 weeks old. The challenge inoculum varied from 10(4) to 10(11) organisms. The disease was characterized by fever, depressed attitude, and decreased appetite. Many calves given larger challenge dose levels also had diarrheic feces containing mucus, fibrin, and blood. Fecal cultures were positive for salmonella. Septicemia occurred in some calves (9 of 15 calves cultured were positive). Eleven calves died and 10 calves survived challenge exposure. Survival was inversely related to the size of the challenge inoculum and directly related (although to a lesser degree) to age of the calf. White blood cell total and differential counts were variable. Both neutropenia and neutrophilia were observed. Plasma proteins decreased markedly in calves with diarrhea, probably indicating fecal protein loss. Fibrinogen increased during the acute stages of diarrhea.     

The Prevalence of Aeromonas Species in Feces of Horses with Diarrhea

Feces collected from 40 horses with diarrhea and 34 horses without diarrhea were examined to determine if an association existed between isolation of Aeromonas spp. and diarrhea. Samples were also examined for Salmonella spp., and identification of viruses and parasite ova. Neither Salmonella spp. nor Aeromonas spp. were isolated from the feces of 34 control horses. Aeromonas spp. were isolated from feces of 22 of 40 (55%) horses with diarrhea. Salmonella spp. were isolated from feces of 8 (20%) horses, and of these, 5 (12.5%) were also positive for Aeromonas spp. Twenty-nine isolates of Aeromonas spp. were recovered from the feces of 22 diarrheic horses. Of these isolates, more than 80% were susceptible on in vitro testing to amikacin, ceftiofur, chloramphenicol, and gentamicin. All isolates were susceptible to enrofloxacin. Diarrheic horses positive for Aeromonas were significantly (P = .04) older than diarrheic horses negative for Aeromonas spp. A significantly greater number of fecal samples were positive for Aeromonas spp. during March through August than samples examined in other months (P = .014). Results of this study indicate that Aeromonas spp. should be considered as a cause of diarrhea in horses.     

Detection of toxin genes in Escherichia coli isolated from normal dogs and dogs with diarrhea.

The etiology of acute, nonviral diarrhea in dogs is poorly understood. Enterotoxigenic and verotoxigenic Escherichia coli are causal agents of diarrhea in humans, pigs, and cattle, but the association of these toxigenic E. coli with diarrhea in dogs has not been explored to a significant extent. In this study, DNA hybridization and PCR amplification were used to identify the frequency with which the genes for E. coli enterotoxins (STap, STb, and LTI) and verotoxins (VT1 and VT2) occur in association with diarrhea in dogs. Genes for VT1 (8.9%), VT2 (22.2%), STa (26.7%), and STb (4.4%) were identified in E. coli cultured from feces of 20 of 45 dogs (44.4%) with diarrhea. Genes for VT2, STa, and STb were not identified in feces from normal dogs. Genes for VT1 were observed in similar proportions in fecal samples from diarrheic (8.9%) and normal (12.3%) dogs. Heat labile enterotoxin (LTI) was not detected in fecal samples from either diarrheic or normal dogs. Our results suggest that heat stable enterotoxins and VT2 may be causally associated with diarrhea in dogs. Dogs appear to be able to carry VT1-producing E. coli without showing overt signs of disease.     

Isolation of bovine coronavirus from feces and nasal swabs of calves with diarrhea.

Fecal and nasal samples were collected from 180 calves with diarrhea and 36 clinically normal co-habitants, and tested for virus using HRT-18 cell cultures derived from human rectal adenocarcinoma. A cytopathic virus was isolated from 5 fecal and 56 nasal samples obtained from diarrheic calves. All calves in which the virus was isolated from diarrheic feces were positive for virus isolation from nasal swabs. The virus was also isolated from the nasal swabs of 10 clinically normal calves that were co-habitants with diarrheic calves. Because they were morphologically similar to coronavirus, agglutinated mouse erythrocytes and serologically identical with the Nebraska calf diarrhea coronavirus, new isolates were identified as bovine coronavirus. The demonstration of viral antigens in nasal epithelial cells by a direct immunofluorescence was in close agreement with the virus isolation in HRT-18 cell cultures. This is the first report on the isolation of bovine coronavirus from newborn calves with diarrhea in Japan. The evidence that the virus was frequently isolated from nasal swabs is of great interest for understanding the pathogenesis of bovine coronavirus infection.     

Evaluation of finishing performance,carcass characteristics,acid-resistant E. coli and total coliforms from steers fed combinations of wet corn gluten feed and steam-flaked corn

Crossbred beef steers (n = 615) were used in a 152-d experiment to compare steam-flaked corn (SFC) diets containing 0, 30, or 60% wet corn gluten feed (WCGF). On d 114 to 118, ruminal and fecal samples were collected from 180 steers and analyzed for pH, VFA, and total and acid-resistant Escherichia coli and coliforms. Acid resistance of E. coli and coliform populations was determined by exposure of the samples for 1 h in pH 2, 4, and 7 citric acid/sodium phosphate buffers. Increasing levels of WCGF linearly decreased total ruminal VFA (P = 0.01) and total fecal VFA (P = 0.06), but linearly increased ruminal and fecal acetate:propionate (P < 0.01) ratio and ruminal and fecal pH (P < 0.05). Feeding increasing WCGF levels resulted in a quadratic response (P < 0.05) with respect to numbers of ruminal E. coli and total coliform populations resistant to pH 4 exposure. Steers fed 30% WCGF had higher (0.7 log units) ruminal E. coli and total coliforms after exposure at pH 4 compared to steers fed 0 or 60% WCGF. Populations of E. coli and total coliforms at pH 2 and 7 were similar for all dietary treatments. Dietary WCGF linearly increased DMI (P = 0.07) and liver abscesses (P = 0.03) and linearly decreased dietary NEg (P = 0.02). Average daily gain and feed efficiencies were greatest when steers were offered 30% WCGF (quadratic, P < 0.05). Dietary manipulations that reduce acid concentrations may not correspond to changes in acid resistance of E. coli and total coliform populations detected in the gastrointestinal tracts of cattle. Moderate levels of WCGF complement SFC finishing diets.     

Detection of Escherichia coli Shiga toxin (stx) and enterotoxin (estA and elt) genes in fecal samples from non-diarrheic and diarrheic greyhounds

Virulence factors responsible for acute diarrhea in greyhounds have not been well established. The objective of this study was to determine if a correlation exists between disease and the presence of the Escherichia coli toxin genes in non-diarrheic and diarrheic greyhound feces. DNA extracted from broth cultures was evaluated for the presence of Shiga toxin and enterotoxin genes and broth samples were evaluated for Shiga toxin and heat-labile enterotoxin. Shiga toxin (stx1 and stx2) and enterotoxin (et and estA) genes were identified in both non-diarrheic and diarrheic samples after in vitro cultured of swabs at 37 degrees C for 16-24h. The stx1 gene was present in 3% of non-diarrheic and 15% diarrheic samples and the stx2 gene was identified in 36 and 23%, non-diarrheic and diarrheic samples, respectively. Shiga toxin was present in 48% diarrheic and 25% of the non-diarrheic in vitro cultured samples. The elt gene was detected in vitro cultured swabs in 12% of the non-diarrheic and 7% of the diarrheic samples. Labile toxin was present in the feces of small numbers of both groups of dogs. A significant correlation existed between the presence of both stx1 genes and Shiga toxin in feces, and lack of disease in non-diarrheic (P=0.01) and presence of disease in diarrheic (P=0.024) greyhounds. Correlation between production of Shiga toxin and detection of stx1 or stx2 was significant in both the diarrheic and non-diarrheic feces (P=0.03); however, only the presence of stx1 correlated with diarrhea in both groups of samples (P<0.008). The incidence of toxigenic E. coli in both non-diarrheic and diarrheic greyhounds indicates a zoonotic potential from dogs to humans and requires further study.     

Fructooligosaccharide supplementation in the yearling horse: effects on fecal pH, microbial content, and volatile fatty acid concentrations

Short-chain fructooligosaccharides (FOS) were supplemented to the diets of nine quarter horses ranging in age from 489 to 539 d with initial BW averaging 400.6 +/- 21.2 kg. The objectives of this study were to determine the effects of dietary FOS on the fecal responses in terms of pH, the microbial population, and VFA concentrations. The horses were used in a 3 x 3 replicated Latin square design, fed according to NRC requirements, and their individual diets were supplemented with no FOS (CON), 8 g of FOS/d (LOW), or 24 g of FOS/d (HIGH) over three 10-d feeding periods. On the last 3 d of each 10-d feeding period, a single fecal sample was collected between 0730 and 0930. Fecal pH decreased linearly (P = 0.01) from 6.48 with the CON diet to 6.38 with the HIGH diet, but there was no change (P = 0.19 for linear effect) in fecal consistency among treatments. A quadratic effect (P < 0.01) was observed for fecal Escherichia coli population, but no difference (P = 0.88 for linear effect) was found in fecal Lactobacilli enumeration among treatments. The presence of fecal Bifidobacteria was unable to be confirmed and was therefore not reported. Fecal acetate concentrations increased linearly (P = 0.03), with means of 2.13, 2.18, and 2.52 mg/g of wet feces for CON, LOW, and HIGH treatments, respectively. Similarly, fecal propionate concentrations increased linearly (P = 0.01), with means of 0.58, 0.64, and 0.73 mg/g for CON, LOW, and HIGH treatments, respectively. Fecal butyrate concentrations also increased linearly (P = 0.02), with means of 0.40, 0.46, and 0.54 mg/g for CON, LOW, and HIGH treatments, respectively. Total VFA (P = 0.01) and lactate (P = 0.02) concentrations increased linearly, with total VFA means of 3.47, 3.69, and 4.25 mg/g for CON, LOW, and HIGH treatments, respectively, and lactate means of 0.36, 0.41, and 0.47 mg/g for CON, LOW, and HIGH treatments, respectively. Supplementing FOS in diets fed to yearling horses altered fecal microbial populations, fecal VFA concentrations, and pH.     

D-lactate production and excretion in diarrheic calves

The origin of D-lactate, the most important acid contributing to metabolic acidosis in the diarrheic calf, is unknown. We hypothesized that because D-lactate is produced only by microbes, gastrointestinal fermentation is the source. The objective of this study was to determine whether D-lactate production occurs in the rumen, colon, or both, and to measure D- and L-lactate concentrations in urine. Fecal, rumen, blood, and urine samples were obtained from 16 diarrheic and 11 healthy calves. Serum electrolyte concentrations were measured in both groups, and blood gas analyses were performed for diarrheic calves. All samples were analyzed for D- and L-lactate by high performance liquid chromatography (HPLC). Diarrheic calves were generally hyperkalemic with high serum anion gap, depressed serum bicarbonate, and low blood pH. L-lactate was markedly higher in rumen contents (22.7 mmol/ L [median]) and feces (8.6 mmol/L) of diarrheic calves than healthy calves (0.5 mmol/L and 5.1 mmol/L, respectively), but not different in serum or urine. Rumen, fecal, serum, and urine D-lactate concentrations were all significantly higher (P < .05) in diarrheic calves (17.0, 25.4, 13.9, and 19.2 mmol/L, respectively) than in healthy calves (0.5, 9.1, 1.4, and 0.5 mmol/L, respectively). Higher D-lactate concentrations in the rumen and feces of diarrheic calves suggests these sites as the source of D-lactate in blood and urine.     

检测绵羊源爱知病毒D型荧光RT-PCR方法的建立和应用

绵羊源爱知病毒D型(ovine Aichivirus D)是在国内绵羊中新发现的病毒,本研究根据绵羊源Aichivirus D 3D基因序列设计检测引物,通过反应体系和条件优化,成功建立了检测绵羊源Aichivirus D的TB Green染料法荧光RT-PCR方法,该方法特异性和稳定性良好,灵敏度高。对2020年4月―2021年5月采集自四川6个场253份绵羊粪便样本(健康羊的133份,腹泻羊的120份)进行检测,结果该病毒的平均检出率为8.7％,场阳性率为66.7％。其中,腹泻粪便样本中绵羊源Aichivirus D阳性率(17.5%)显著高于非腹泻粪便样本中绵羊源Aichivirus D阳性率(0.75%,P<0.001)。表明绵羊源Aichivirus D可能是引起以上地区绵羊腹泻的病原。从绵羊源Aichivirus D的阳性样本中成功克隆出大小为1 422 bp的13个完整的绵羊源Aichivirus D 3D基因,其核苷酸相似性为99.4％～100.0％。遗传演化分析发现这13株绵羊源Aichivirus D 3D基因与本实验室前期研究上传的绵羊源Aichivirus D 3D基因共同聚为单独的一个大支。本研究为绵羊源Aichivirus D的分子检测提供了一种新的方法和基础流行病学数据。     

Detection and molecular characterization of porcine group C rotaviruses in South Korea

Group C rotaviruses (GCRVs) cause acute diarrhea in humans and animals worldwide and the evidence for a possible zoonotic role of GCRVs has been recently provided. However, there is little evidence of porcine GCRV infections or of their genetic diversity in South Korea. We examined 137 diarrheic fecal specimens from 55 farms collected from six provinces. RT-PCR utilizing primer pairs specific for the GCRV VP6 gene detected GCRV-positive reactions in 36 (26.2%) diarrheic fecal samples. Of these, 17 samples (12.4%) tested positive for porcine GCRVs alone and 19 samples (13.8%) were also positive for other pathogens. Other enteric pathogens except for GCRV were detected in 64 feces samples (46.7%) and no enteric pathogens were evident in 37 feces samples (27.0%). Phylogenetic and sequence homology analyses of GCRV partial VP6 gene between 23 Korean and other known porcine GCRVs demonstrated that Korean strains belonged to the porcine lineage. Furthermore, one Korean porcine strain shared the highest nucleotide (89.7–89.0%) and deduced amino acid sequence (92.9–93.9%) identities with bovine GCRV strains and was placed in the bovine GCRV lineage indicative of bovine origin. In conclusion, porcine GCRV infections are widespread in piglets with diarrhea in South Korea. The infecting porcine GCRVs mostly belong to the porcine lineage with the exception of one bovine-like GCRV, which possibly originated from bovine GCRV due to interspecies transmission.     

Optimization of a species-specific polymerase chain reaction assay for identification of Pentatrichomonas hominis in canine fecal specimens

OBJECTIVE: To determine the optimum reaction conditions and detection limits of PCR assay for identification of Pentatrichomonas hominis in DNA extracted from canine feces. SAMPLE POPULATION: DNA extracted from feces of 4 dogs with diarrhea from which trichomonads were observed, 81 dogs that had feces submitted to a diagnostic laboratory, and 19 dogs residing in a laboratory animal facility. PROCEDURES: Optimum reaction conditions and absolute and practical detection limits of 2 P hominis 18S species-specific primer pairs were determined by use of an in vitro cultivated canine isolate of P hominis in the presence and absence of canine feces. The optimized PCR assay was applied to amplification of P hominis 18S rRNA genes from DNA extracted from the feces of dogs. RESULTS: Under optimized conditions, a primer pair was identified as able to detect as few as 1 P hominis organism/180-mg fecal sample. The PCR assay identified P hominis in diarrheic feces of 4 dogs in which trichomonads were seen by light microscopy. The P hominis genes were not amplified from other fecal samples examined. CONCLUSIONS AND CLINICAL RELEVANCE: Molecular identification of P hominis in feces of 4 dogs with trichomonosis and diarrhea reported here validates the identity of this species in such infections. Sensitive and specific PCR amplification of P hominis 18S rRNA genes from DNA extracted from feces will directly facilitate studies examining pathogenicity of this trichomonad and enable differentiation of P hominis from other known or novel species of trichomonads that may infect the gastrointestinal tract of dogs.