Prevalence of Cryptosporidium, Giardia and Eimeria infections in post-weaned and adult cattle on three Maryland farms

The prevalence of Cryptosporidium, Giardia and Eimeria, in healthy, asymptomatic, post-weaned and mature cattle was investigated on three Maryland farms. One farm, a dairy research facility, had 150 multiparous Holstein milking cows; 24 were examined and Cryptosporidium andersoni was detected in three (12.5%) but neither Giardia nor Eimeria was detected. The second farm, a commercial dairy, had 57 multiparous Holstein milking cows and an equal number of heifers. Of 19 cows examined, C. parvum, Giardia duodenalis, and Eimeria bovis and/or E. ellipsoidalis were detected in two (10.5%), two (10.5%) and one (5.26%) cow, respectively. Of 23 heifers examined, C. parvum, Giardia, and E. bovis and E. ellipsoidalis, was detected in two (8.7%), four (17.4%), and five (21.7%), heifers, respectively. The third farm, a beef cattle breeding and genetics research facility, had 180 7- to 9-month old purebred black Angus. Of 118 examined for C. parvum and Giardia, 34 (28.8%) and 44 (37.3%) were positive, respectively, of 97 examined for E. bovis and/or E. ellipsoidalis 32 (33.0%) were positive. These findings, based on a method with a minimum detection level of 100 oocysts of C. parvum/g of feces, which underestimates the number of infected cattle, clearly demonstrate the presence of low level, asymptomatic infections in post-weaned and adult cattle in the United States and indicate the potential role of such cattle as reservoirs of infectious parasites.     

Prevalence and pathogenicity of Cryptosporidium andersoni in one herd of beef cattle

Over a 35-week period from January to July 2002, a breed of Hereford beef cattle (H) and their hybrids were monitored. Five hundred and ninety-nine individual fecal samples from calves and 96 samples from their mothers were examined. First excretion of Cryptosporidium andersoni oocysts in calves was found in the 9th week after the start of calving (a calf 63-day old). The prevalence of C. andersoni in calves ranged from 11.1 to 92.9% depending on age. The mean prevalence in their mothers was found to be 43.8%. The size of oocysts was 8.48 +/- 0.78 x 6.41 +/- 0.59 microm. Infection intensity in calves ranged from 32 000 to 4 375 000 oocysts per gram (OPG) and in mothers from 78 000 to 2 552 000 OPG. Three cases of abomasal cryptosporidiosis slaughtered at the age of 81, 157 and 236 days were examined histologically and ultrastructurally [transmission electron microscopy (TEM) and scanning electron microscopy (SEM)]. Cryptosporidium infection of the abomasum was located in the upper half of the mucosal glands in the plicae spirales of the fundus, corpus and near the ostium omasoabomasicum. Cryptosporidia were not located in the glandular epithelium of the pars pylorica in the abomasum minimally 10 cm from pylorus. Histopathological changes in the site of cryptosporidial infection in the abomasum had a non-inflammatory character and included distinctive dilatation of infected parts of the glands with atrophy and metaplasia of the glandular epithelial cells, goblet cell activation and mucus hyperproduction. The TEM revealed a relatively small number of Cryptosporidium life cycle stages attached to glandular epithelial cells. In SEM the inner mucosal abomasal surface appeared swollen but was never infected by cryptosporidia.     

屠宰场猪链球菌分离菌株分型及其致病性和耐药性分析

为了解吉林省猪链球菌的流行情况,从屠宰场采集的猪咽拭子和鼻拭子样品中分离鉴定猪链球菌,并进行菌株血清型、基因型和毒力表型的鉴定,以及致病性和耐药性的分析。结果表明,从100份样品中共分离鉴定猪链球菌104株,其中29株鉴定为血清2型、9型和1型等几种常见的致病性血清型,其他75株不属于常见的致病性血清型。血清2型的菌株中,2株经鉴定为ST1基因型和mrp+epf+sly+毒力型,并经动物试验鉴定为强毒菌株;其他菌株均为ST28基因型和mrp+epf-sly-毒力型,具有中等毒力。血清9型的菌株,均为mrp-epf-sly-毒力型,动物试验鉴定均为强毒菌株。根据Kirby-Bauer纸片扩散法的药敏试验结果,98%的猪链球菌分离株对四环素耐药;对大环内酯类、克林霉素和链霉素的耐药率都在50%以上;对β-内酰胺类、氯霉素和喹诺酮类抗生素的耐药率小于20%。总体分析,从屠宰场分离的猪链球菌强毒菌株所占的比例并不高,但是菌株多重耐药的情况非常严重。     

Prevalence of Cryptosporidium spp. in pigs in Shanghai, China

A survey on prevalence of Cryptosporidium spp. in pigs at 12 farms in 8 suburban districts and 1 county of Shanghai was conducted under Sheather's sucrose flotation protocol and modified acid-fast stain methods from 2006 to 2009. A total of 2323 faecal samples were collected and Cryptosporidium spp. oocysts were detected in 800 samples (34.4%). Cryptosporidium was found in all 12 pig farms. Significant variations of prevalence were observed in different farms ranging from 14.1% to 90.6%. A follow-up survey on a positive pig farm for 13 consecutive months revealed that the most serious infection of Cryptosporidium spp. in pigs happened in winter and spring, and the lowest infection season was summer. Cryptosporidium spp. infection was mainly found in piglets within 2 months and no infection was found among those pigs of 90-180 days of age. The genotype analyses were carried out through PCR-RFLP and partial sequences analysis of small subunit ribosomal RNA (SSU rRNA) in some of the positive samples. Cryptosporidium suis (57/69, 82.6%), Cryptosporidium pig genotype II (6/69, 8.7%) and mixed infection of above two species/genotype (6/69, 8.7%) were found to be the main species/genotype in pigs in Shanghai area.     

猪链球菌扁桃体分离株的毒力因子分布特征与致病性

本研究设计并合成7对引物,用PCR方法对猪链球菌7种主要毒力因子,包括谷氨酸脱氢酶(gdh)、溶血素(sly)、胞外蛋白因子(ef)、溶茵酶释放蛋白(mrp)、纤连蛋白/血纤蛋白原结合蛋白(fpbs)、三磷酸甘油醛脱氢酶(gadph)和毒力相关序列orf2进行检测,分析了29株猪链球菌扁桃体分离株的毒力因子分布特征.在被检的25株2型菌株中,共检测出7个基因型.其中10株(40%)的基因型为cps2/gdh+/sly+/ef+/mrp+/fbps+/gadph+/orf2+,7株(28%)表现为cps2/gdh+/sly-/ef-/mrp+/fbps+/gadph+/orf2+,4株(16%)表现为eps2/gdh+/sly-/ef+/mrp+/fbps+/gadph+/orl2+,基因型表现为eps2/gdh+/sly+/ef+/mrp+/fbps+/gadph+/orf2-、cps2/gdh+/sly-/ef-/mrp+/fbps+/gadph+/orf2-、eps2/gdh+/sly-/ef-/mrp*/fbps+/gadph+/orf2+、eps2/gdh+/sly-/ef-/mrp-/fbps-/gadph+/orf2-各1株;1株猪链球菌7型(SS7)分离株,基因型表现为cps7/gdh+/sly+/ef-/mrp-/fbps+/gadph+/orf2+;3株猪链球茵9型(SS9)扁桃体分离株,均表现为cps9/gdh+/sly-/ef-/mrp-/fbps+/gadph+/orf2+.可见我国SS分离株的毒力基因分布较为复杂,而且SS2的优势流行菌株是同时具有多种毒力因子的高致病茵株.通过对不同毒力基因型毒株对西藏小型猪的感染试验发现:毒力基因型为cps2/gdh+/sly-/ef+/mrp+/fbps+/gadph+/orf2+的健康扁桃体分离株SH06-21D对西藏小型猪有较强的致病性,仅次于标准株HA9801,而基因型为sly-/ef-/mrp-/fbps-/gadph+/orf2-的健康扁桃体分离株GZ06-122B对西藏小型猪不表现明显的致病性.     

Prevalence and Pathogenicity of Cryptosporidium suis in Pre‐ and Post‐weaned Pigs

A total of 4338 faecal samples, 135 of sows, 3368 of pre‐weaned and 835 of post‐weaned piglets from eight farms in South Bohemia, Czech Republic were collected and examined for Cryptosporidium infection. No sow, but 5.7% pre‐weaned and 24.1% post‐weaned piglets were positive for Cryptosporidium infection. No relationship was found between diarrhoea and Cryptosporidium infection in any of the different age groups (pre‐ and post‐weaned piglets). Four piglets, which were sporadically shedding cryptosporidia in faeces, were necropsied. Neither clinical signs of diarrhoea nor macroscopical changes were found. Histologically, a moderate infection of cryptosporidia was detected in the glandular epithelium along the large intestine, with predisposition to the ansa centralis of the colon. No inflammatory response in the lamina propria was observed. Cryptosporidia were also commonly found in the glandular epithelium of submucosal lymphoglandular complexes in the colon. Cryptosporidium isolates from all farms were identified as Cryptosporidium suis using molecular markers (SSU rRNA). All of the C. suis strains obtained were larger [6.2 (6.0–6.8) × 5.5 (5.3–5.7) μm] than any isolate described so far [4.6 (4.4–4.9) × 4.2 (4.0–4.3) μm] and did not appear to be infective for neonatal BALB/c mice.     

Prevalence and genotypes of Giardia duodenalis in post-weaned dairy calves

To determine the prevalence of Giardia genotypes in post-weaned dairy calves, fecal specimens were collected from 3 to 11-month-old dairy calves per farm on two farms in Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida. Specimens cleaned of fecal debris and concentrated using CsCl density gradient centrifugation were stained and examined by immunofluorescence microscopy and also subjected to PCR and DNA sequence analysis. Overall, PCR provided more sensitive detection than IFA. Prevalence of Giardia infection, as detected by PCR ranged from 20% on NC-2 to 81% on VT-2, with an overall prevalence of 52% (237 positive samples out of 456 total). DNA sequence analysis of the 16S rRNA gene revealed 87% of the 237 Giardia isolates were Assemblage E, and 13% were Assemblage A although the prevalence of these genotypes varied greatly from farm to farm, with five farms having no Assemblage A Giardia. Therefore, Assemblage E was present in 45% of all animals tested and Assemblage A was present in 7% of the animals. Thus, while many of the calves were infected with a genotype that is not known to be infectious for humans, post-weaned calves on nine of 14 farms did harbor Assemblage A Giardia. Therefore calves should be considered as a potential source of human infectious cysts in the environment, with some farms representing a much higher risk than others.     

Prevalence and risk factors associated with Cryptosporidium oocysts shedding in pigs in Central Vietnam

We investigated the prevalence and risk factors associated with Cryptosporidium oocysts shedding in pigs in Central Vietnam. A total of 740 single fecal samples collected from diarrheic and non-diarrheic pigs on 89 farms were screened by the modified Ziehl-Neelsen staining method. Prevalence at the animal and the farm levels were 18.1% (134/740) and 71.9% (64/89), respectively. Risk factors for the infection were identified using univariate and multivariate logistic regression analysis. The results revealed that age, sanitary condition and topography were significantly associated with oocyst shedding (P<0.05). Pre-weaned piglets were at the highest risk for infection, followed by post-weaners, sows and finishing pigs. Good sanitary conditions showed positive effects in decreasing oocysts shedding. Topographically, Cryptosporidium was more common in mountainous zone than that in coastal delta zone. There was an association between the occurrence of diarrhea and the level of Cryptosporidium oocyst excretion within infected pigs. This is the first epidemiological investigation of prevalence and risk factors of Cryptosporidium in pigs in Vietnam.     

Prevalence of Streptococcus suis types 1 and 2 in domestic pigs in Australia and New Zealand

Using an indirect fluorescent antibody test, 54 per cent of 734 palatine tonsils of conventional pigs slaughtered in Australia and New Zealand were found to be infected with Streptococcus suis type 1 and 73 per cent of 959 were infected with S suis type 2. Variations in the prevalence of infection in pigs from different herds were thought to be due to differences in the sample sizes rather than to real differences in the prevalence between herds. The prevalence of infection with S suis was similar in pigs of either sex and in different age groups. Streptococcus suis type 2 was detected in the blood of 3 per cent of apparently normal pigs slaughtered at a meat processing plant. The presence of this organism in edible tissue may pose a health risk to consumers and meat-workers. Both S suis types 1 and 2 were detected in the vaginas and uteri of slaughtered pigs and the female reproductive tract could be another site for the carriage of infection. Piglets from sows with vaginas infected with S suis type 2 became infected earlier than piglets from sows with uninfected vaginas. No infected male reproductive tracts were detected and venereal transmission of S suis therefore appears unlikely. Three specific pathogen free herds were found to be free from infection with both S suis types 1 and 2. It is concluded that hysterectomy derived piglets are delivered free from infection, whereas some piglets born to sows with uterine and vaginal infections are either born infected or become infected at, or soon after, birth.     

Prevalence of the suilysin gene in Streptococcus suis strains isolated from diseased and healthy carrier pigs

Knowledge of virulence factors of Streptococcus suis is limited. Several virulence factor candidates have been proposed, among them suilysin, which is responsible for a toxic effect on epithelial cells. The aim of this study was to detect the suilysin gene sequence in Streptococcus suis strains of various origin. In total 63 Streptococcus suis isolates were investigated. Forty four of them originated from tissues of streptococcosis affected animals. The remaining 19 strains were isolated from tonsils of healthy carrier pigs. Suilysin gene specific sequence was detected in 79% of the strains tested. In isolates obtained from pigs with signs of streptococcosis this gene sequence was recorded in 85% of cases. In Streptococcus suis strains isolated from healthy carrier pigs the suilysin gene was detected in 63% of the isolates. It seems that suilysin toxic activity is only one of the many steps involved in the pathogenesis of Streptococcus suis infection and that strain's virulence cannot be stated only on the basis of suilysin gene sequence presence.     

猪附红细胞体病原特征及其致病性分析

为研究猪附红细胞体的病原形态学特征及其致病特性,给6只临床健康小鼠成功感染了猪附红细胞体病,对试验组小鼠感染后的不同时期采集的血样进行了鲜血压片、姬姆萨染色及扫描电子显微镜下的形态学检查,红细胞计数和以P1:5'-GCATGCCCAGTCCCCAAGGA-3'和P2:5'-TGCGGGGAGTACGTGGGAAGG-3'为特异性引物,利用2003年Ludwig E报道的方法对待检血样中附红细胞体DNA的PCR鉴定及测序.1 000倍油镜下观察到单个或连成线状的附红细胞体附着于红细胞表面,红细胞变成刺状、菠萝状或"口"字形等不规则形状;12 000倍扫描电镜下观察到了"逗点"状及以纤丝与红细胞膜相连的附红细胞体;红细胞计数结果显示:与对照组相比试验组小鼠在接种后第7、10、14天出现大量异型性红细胞,其中第10天严重变形红细胞数量最多.说明猪附红细胞体病发病明显期在感染后第7～14天,14 d以后逐渐消散;PCR扩增得到782 bp的DNA片段,经序列分析与序列号为AJ504999的相应片段同源性为99.74%.本试验为猪附红细胞体的病原特性的研究提供了科学依据,为猪附红细胞体病发病机理等方面更深入的研究奠定了基础.     

Prevalence, capsular type and antimicrobial susceptibility of Streptococcus suis isolated from slaughter pigs in Korea.

This study was undertaken to determine the prevalence, capsular serotype, and antimicrobial susceptibility of Streptococcus suis isolated from slaughter pigs. Capsular serotype and antimicrobial susceptibility were determined by coagglutination test and agar dilution minimum inhibitory concentration, respectively. Streptococcus suis was isolated from 55 of the 406 palatine tonsillar samples tested (13.8%) and 14 of the 29 sampled herds (48.3%). Of the 55 isolates recovered from slaughter pigs, 26 (47.3%) were untypeable. Of the remaining 29 isolates, capsular serotypes 9 (9 isolates) and 16 (4 isolates) were the most common, followed by capsular serotypes 4 (3 isolates) and 7 (3 isolates). Every capsulated isolate was typeable and no palatine tonsillar sample yielded more than one serotype. Most of isolates were susceptible to low concentrations (MIC90) of amoxicillin (2 microg/mL), ceftiofur (1 microg/mL), and penicillin (1 microg/mL). No correlation was found between antimicrobial susceptibility and capsular serotype.     

Prevalence of Mycoplasma suis in slaughter pigs, with correlation of PCR results to hematological findings

Porcine infectious anemia is a well-known disease that occurs worldwide and is caused by the unculturable hemotrophic bacterium Mycoplasma suis. The actual prevalence and impact of M. suis infections, however, remain fairly unknown. This study examined the prevalence of M. suis in post-weaning pigs by employing a quantitative real-time LightCycler PCR. M. suis infections were detected in 164 out of 1176 feeder pigs (20-30 kg; 13.9%) as well as on 79 out of 196 pig farms (40.3%). The comparison of PCR results with microscopic investigation of acridine-orange-stained blood smears revealed a considerably lower sensitivity of the microscopic method: only 35 out of 1176 blood smears were microscopically positive. The microscopic detection of M. suis was shown to be closely linked to the bacterial load in the blood. M. suis infection is associated with significantly decreased hematocrit, erythrocyte counts and hemoglobin concentrations as well as significantly higher bilirubin concentrations. Furthermore, M. suis blood loads were significantly associated with erythrocyte count, hematocrit, hemoglobin, glucose and iron concentrations indicating that high M. suis loads are connected with clinical anemia. In conclusion, this study has shown, that M. suis infections are often under-diagnosed in pig husbandry and can therefore lead to considerable economic profit losses in pig husbandry. Furthermore, our study has shown that the LightCycler PCR could be an appropriate tool for a sufficiently coherent identification of M. suis in latent carrier animals in view of introducing effective treatment and disease control measures.     

Actinobacillus suis infection in pigs in Australia

This is the first report of the isolation of Actinobacillus suis in association with significant disease among preweaned pigs in Australia. Sudden deaths occurred in preweaned piglets at one facility and enlarged joints, particularly the stifles and tarsi, occurred at another. Isolates with the biochemical phenotype and apx genotype profile of A suis were cultured from affected piglets. Both facilities were of high health status and one had undergone a recent depopulation, disinfection and sow repopulation process. Reviewing initial reports of A suis disease overseas, it is apparent that outbreaks occurred sporadically in various locations, with disease occurring among the herds involved for some months only, before development of immunity. A suis disease is now considered an emerging disease on high health status farms.     

Molecular and biological characterisation of Cryptosporidium in pigs

OBJECTIVE: Genetic and biological characterisation of 12 isolates of Cryptosporidium from pigs and comparing them with Cryptosporidium isolates from humans and cattle. DESIGN: Cryptosporidium isolates from pigs were compared with those obtained from human and cattle using rDNA sequence analysis. The infectivity of two of the porcine isolates was determined in neonatal mice and the clinical history of the infected pigs recorded. RESULTS: Pig-derived isolates of Cryptosporidium exhibited two distinct genotypes; a porcine genotype and a bovine genotype, which is common to cattle and other livestock. The porcine genotype did not produce any infection in neonatal mice whereas the bovine genotype did. CONCLUSION: Two distinct genetically and biologically differing strains of Cryptosporidium appeared to be associated with acute diarrhoea in pigs. Whether Cryptosporidium was a primary or secondary pathogen is unclear but warrants further investigation. As the bovine genotype is known to infect humans, the results suggest that pigs can act as reservoirs of cryptosporidial infections for humans and other live-stock. The zoonotic potential of the pig-adapted genotype is uncertain and requires further study.     

Complex epidemiology and zoonotic potential for Cryptosporidium suis in rural Madagascar

Cryptosporidium spp. is the most important parasitic diarrheal agent in the world, is among the top four causes of moderate-to-severe diarrheal disease in young children in developing nations, and is problematic as an opportunistic co-infection with HIV. In addition, Cryptosporidium is a persistent challenge for livestock production. Despite its zoonotic potential, few studies have examined the ecology and epidemiology of this pathogen in rural systems characterized by high rates of overlap among humans, domesticated animals, and wildlife. To improve our understanding of the zoonotic potential of Cryptosporidium species in the rural tropics, we screened humans, livestock, peridomestic rodents, and wildlife using PCR-RFLP and sequencing-based approaches to distinguish species of Cryptosporidium in rural southeastern Madagascar. Cryptosporidium of multiple species/genotypes were apparent in this study system. Interestingly, C. suis was the dominant species of Cryptosporidium in the region, infecting humans (n = 1), cattle (n =  18), pigs (n= 3), and rodents (n = 1). The broad species range of C. suis and the lack of common cattle Cryptosporidium species (Cryptosporidium parvum and Cryptosporidium andersoni) in this system are unique. This report represents the fifth confirmed case of C. suis infection in humans, and the first case in Africa. Few rural human and livestock populations have been screened for Cryptosporidium using genus-specific genotyping methods. Consequently, C. suis may be more widespread in human and cattle populations than previously believed.     

Streptococcus suis isolated from pigs in Finland

A total of 58 Streptococcus suis strains were isolated from deceased pigs submitted to the National Veterinary Institute, Regional Laboratory in Kuopio, Finland, over a 3 1/2 year period, most frequently from cases of pneumonia. The bacteria were isolated from cases of meningitis, sepsis, rhinitis, endocarditis and abortion. S. suis was also isolated from nasal cavity, lung and brain of some sick piglets without signs of inflammation. Further S. suis was detected in 12 out of 107 tonsils of healthy fatteners tested. S. suis strains were identified by biochemical methods followed by typing. The most common capsular types were 7, 3 and 2, respectively. Only one type 1 strain and no types 6 and 9 strains were found. All S. suis strains tested were sensitive to penicillin and ampicillin.S. suis is not uncommon in Finnish pig herds. S. suis may be regarded as a potentially pathogenic organism which under certain predisposing conditions may cause serious disease.     

New view on the age-specificity of pig Cryptosporidium by species-specific primers for distinguishing Cryptosporidium suis and Cryptosporidium pig genotype II

Two species of Cryptosporidium are commonly identified in pigs: Cryptosporidium suis and Cryptosporidium pig genotype II. Detection of Cryptosporidium spp. is routinely based on molecular methods such as polymerase chain reaction (PCR) and subsequent restriction fragment length polymorphism (RFLP) or gene sequencing. However, most of these methods are hampered by low sensitivity to mixed infections. As a solution of this problem, novel species-specific primers were designed and tested in the present study. Sensitivity of our primers was identical to genus-specific primers, but more (1:48) mixed infections were detected using these species-specific primers on 477 DNA samples originating from naturally infected pigs of different age categories. Our results show differences in age-dependent susceptibility of pigs to the infection. Whereas C. suis was found in all tested categories of pigs (1-12 week of age and sows), Cryptosporidium pig genotype II was recorded only in animals older than 6 week of age. Usage of species-specific primers could help to better the understanding of epidemiology of pig specific Cryptosporidium spp. and its occurrence, which, on the basis of our results, is underestimated.