U.S.-type Babesia microti isolated from small wild mammals in Eastern Hokkaido, Japan

Our previous report demonstrated that small wild rodents in Japan harbored two types of novel Babesia microti-like parasites (Kobe and Hobetsu types), but not the type widely distributed throughout the temperate zones of North American and Eurasian Continents (U.S. type). In this study, we surveyed small wild mammals collected at various places in the northern part of Japan, seeking for U.S.-type B. microti. A total of 197 small mammals comprising 10 species, Apodemus speciosus, A. argenteus, Clethrionomys rufocanus, C. rutilus, Eothenomys andersoni, Microtus montebelli, Tamias sibiricus, Sorex unguiculatus, S. caecutiens, and Urotrichus talpoides, were examined. Babesia parasites were detected in A. speciosus, C. rufocanus, C. rutilus, M. montebelli, S. unguiculatus, and S. caecutiens by microscopy of blood smears and by PCR targeting babesial nuclear small-subunit rRNA (rDNA) and beta-tubulin genes. Inoculation of their bloods into experimental animals gave rise to 23 parasite isolates, which included 16 from A. speciosus, 4 from C. rufocanus, and 1 each from C. rutilus, M. montebelli and S. unguiculatus. Sequencing analyses of their rDNA and beta-tubulin genes revealed that, of the 23 isolates, 20 and 3 were of Hobetsu and U.S. types, respectively. The U.S.-type B. microti strains isolated in Japan, however, were distinguishable from the isolates in the United States when their beta-tubulin gene sequences and antigen profiles in Western blots were compared. We conclude that U.S.-type B. microti exists in Japan although it has been genetically and antigenically diversified from that distributed in the United States. The results also suggest that not only rodents, but also some insectivores may serve as reservoirs for the agent of human babesiosis.     

Anisakidae nematodes and Trypanorhyncha cestodes of hygienic importance infecting the king mackerel Scomberomorus cavalla (Osteichthyes: Scombridae) in Brazil

From February to October 2007, thirty specimens of the king mackerel, Scomberomorus cavalla (Cuvier, 1829) were purchased from markets in the municipalities of Niterói and Rio de Janeiro. The fishes were measured, filleted and further had their organs investigated for helminths. Ten out of the thirty fish specimens were parasitized with anisakid nematodes represented by Anisakis sp. and Contracaecum sp. with prevalence of 1% and 16%, mean intensity of 2 and 3.31 and mean abundance of 0.02 and 0.53, respectively. The infection range with Contracaecum sp. was 1-9. The sites of infection were the stomach serosa and mesentery. Seventeen fish specimens (53%) out of the 30 investigated were parasitized with Trypanorhyncha metacestodes, identified as Callitetrarhynchus gracilis, Pterobothrium crassicole, Callitetrarhynchus speciosus and Tentacularia coryphaenae in the mesentery, with prevalence of 26, 20, 6, 3%, intensity and mean intensity of 3.25, 3.5, 1, 2 and mean abundance of 0.86, 0.7, 0.06 and 0.06, respectively. The infection range due to C. gracilis and P. crassicole were of 1-5 and 1-20, respectively. Anisakis sp., C. speciosus and P. crassicole are reported in S. cavalla for the first time. Considerations on the zoonotic potential of the parasites and their rules in sanitary inspection are presented.     

Detection of Babesia microti-like parasite in filter paper-absorbed blood of wild rodents

The first case of human babesiosis was reported in Japan. The epidemiology of this disease in Japanese nature remains unclear. In this study, 97 common field mice captured in Hokkaido, Japan, were examined. Blood specimens absorbed onto filter papers were eluted and tested by nested PCR using specific primers for the B. microti nuclear small subunit rRNA genome. Twenty-three percent (11/47) of Apodemus speciosus and four percent (2/50) of Clethrionomys rufocanus were positive. The 159-bp primary sequences of PCR products tested exhibited 97.5% and 96.8% homology with those of the human isolate in Japan and of U.S. strains of B. microti, respectively.     

Antimicrobial susceptibility of Escherichia coli isolates from wild mice in a forest of a natural park in Hokkaido, Japan

To reveal the antimicrobial susceptibilities of Escherichia coli isolates from wild mice, 81 E. coli isolates were obtained from 109 voles (Clethrionomys spp.), 52 large Japanese field mice (Apodemus speciosus) and 19 small Japanese field mice (A. argenteus) captured in a forest of a natural park in Hokkaido, Japan. Seventy-eight of the 81 E. coli isolates were susceptible to all 10 antimicrobial agents tested. One E. coli isolate was resistant to ampicillin, dihydrostreptomycin, kanamycin, chloramphenicol and oxytetracycline. Two isolates were resistant to oxytetracycline. A low prevalence of antimicrobial resistance was maintained among wild mice that inhabited the forest.     

Detection of Cryptosporidium muris type oocysts from beef cattle in a farm and from domestic and wild animals in and around the farm

Cryptosporidium muris type oocysts were detected from 21 of 516 beef cattle in a farm. Then we surveyed Cryptosporidium oocysts in 348 beef and dairy cattle, 500 pigs, 101 dogs, 38 wild animals and 11 zoo-kept animals in and around the farm. Oocysts were detected from only 2 of 25 Japanese field mice, Apodemus speciosus in the same farm. Gene analysis suggested that the oocysts were different from the C. muris type bovine isolate.     

Clinical problems of sloths (Bradypus sp. and Choloepus sp.) in captivity.

A 20-yr retrospective study of disease prevalence was carried out for 51 sloths (34 Bradypus sp. and 17 Choloepus sp.) at the S?o Paulo Zoo. A total of 81 clinical disorders were detected, including nutritional (45.7%), digestive (12.3%), and respiratory (12.3%) problems and injuries (6.1%). A definitive diagnosis was not possible in 8.6% of the cases. The incidence of disease varied according to seasonal climate (winter, 32.5%; spring, 24%; summer, 22.9%; autumn, 20.5%), time in captivity (96.4% of diseases occurred within the first 6 mo and 3.6% occurred thereafter), and type of enclosure (quarantine cage, 96.4%; exhibition enclosure, 3.6%). Both young animals (86.7%) and adults (3.2%) were affected. Parasites were identified by fecal examination in 45.4% of animals with clinical illness (Ascaris sp., 80%; Coccidia sp., 20%). Bacteria such as Salmonella enteritidis, Escherichia coli, and Citrobacter freundii were isolated from feces and/or organs. The first 6 mo in captivity are critical for these animals. Proper management and early identification of medical conditions in captivity have implications for sloth population in the wild.     

长角血蜱肠道细菌的分离与鉴定

为深入研究肠道细菌在蜱的生理学和传播病原上的作用,本研究进行了长角血蜱肠道细菌的分离鉴定。按照细菌传统分离方法并结合16S rDNA测序鉴定,获得7个分离株,这些菌株分别属于显核菌属(Caryophanon sp.)、考克氏菌属(Kocuria sp.)、葡萄球菌属(Staphylococcus sp.)、微杆菌属(Microbacterium sp.)和短杆菌属(Brevibacterium sp.)5个菌属。     

Molecular detection of an Ehrlichia-like agent in rainbow trout (Oncorhynchus mykiss) from Northern California

Ehrlichia DNA was identified by nested PCR in rainbow trout (Oncorhynchus mykiss) collected from a creek in northern California where Potomac horse fever is endemic. Ehrlichia DNA was found in tissues from several organs including the gills, heart, spleen, liver, kidneys and intestine of trout and from three different adult digenetic trematodes (Deropegus sp., Crepidostomum sp., Creptotrema sp.) parasitizing the gallbladder and/or the intestine of the trout. Sequencing of PCR-amplified DNA from the 16S rRNA gene indicated that the source organism was most closely related to the sequences of E. risticii (level of sequence similarity 96.0%), the SF agent (95.9%), E. sennetsu (95.8%), and Neorickettsia helminthoeca (95.3%). The data suggest that trout and parasitic trematodes may be involved in the epidemiology of an Ehrlichia-like agent belonging to the E. sennetsu genogroup. Whether the fish agent infects horses, dogs, or human beings, and whether it causes disease, remain to be determined.     

Oribatid mites (Acari, Oribatida) as intermediate hosts of tapeworms of the Family Anoplocephalidae (Cestoda) and the transmission of Moniezia expansa cysticercoids in South Africa

Six species of adult oribatid mites (Galumna racilis, Kilimabates pilosus, Kilimabates sp., Scheloribates fusifer, Muliercula ngoyensis and Zygoribatula undulata) and two immature stages belonging to the superfamilies Galumnoidea and Ceratozetoidea were isolated from a lawn (mixed Pennisetum and Cynodon spp.) at Onderstepoort Veterinary Institute, South Africa. The mites were subsequently used in an infection trial using Moniezia expansa eggs. Tapeworm cysticercoids were recovered in G. racilis, K. pilosus, Kilimabates sp., S. fusifer, M. ngoyensis and Z. undulata, as well as in immatures of Ceratozetoidea.The percentage of infected mites was 7,6, 6,3, 16,4, 66,7, 57,1, 60,0 and 46,7% respectively. Immatures of Galumnoidea did not become infected. The highest number of cysticercoids isolated from one individual was six from an adult S. fusifer.     

Helminth parasites in faecal samples from the endangered Iberian lynx (Lynx pardinus)

The Iberian lynx is the most endangered felid in the world. Enteropathogens may threaten its survival, and therefore we analysed faecal samples from 66 different individuals (37 males and 29 females), the largest population representation studied to date. The samples were obtained from November 2005 to October 2008 in the two areas where the Iberian lynx survives: Sierra Morena and Do?ana (Andalusia, southern Spain). A total of 56.1% samples were parasitized with at least 6 species of helminths, including two cestodes (Hymenolepis spp. and Taenia spp.) and four Nematodes (Ancylostoma spp., Toxocara spp., Toxascaris leonina, and Capillaria sp.). In this work, the presence of Hymenolepis is reported for the first time in Lynx pardinus. The relevance of our findings is discussed focussed on the conservation of this endangered felid.     

Gastrointestinal parasites of mountain gorillas (Gorilla gorilla beringei) in the Parc National des Volcans, Rwanda.

Ninety-eight fecal samples were collected from 74 free-living mountain gorillas (Gorilla gorilla beringei) from the Parc National des Volcans, Rwanda, between July 1995 and January 1997 and examined for parasites by Sheather's sugar and zinc sulfate flotation methods, trichrome staining, and larval cultures. All samples contained at least one parasite. Seventeen endoparasites were identified, including eight protozoa, seven nematodes, one cestode, and one trematode. Two species of arthropod mite were also recovered from the fecal samples. Parasites observed on fecal examinations included strongyle/trichostrongyle-type eggs (72/74) (representing Oesphagostomum sp., Trichostrongylus sp., Hyostrongylus spp., and possibly Murshidia sp.), Strongyloides sp. (1/74), Trichuris trichiura (2/74), Probstmayria sp. (7/74), Anoplocephala sp. (63/74), Entamoeba hartmanni cysts and trophozoites (19/70), Endolimax nana cysts (31/70), Iodamoeba buetschlii cysts (11/70), Endolimax nana or Iodamoeba buetschlii trophozoites (63/70). Entamoeba coli cysts and trophozoites (14/70), Entamoeba histolytica trophozoite (1/70), Chilomastix sp. cysts and trophozoites (31/70), and Giardia sp. cysts (2/70). In addition, one ascarid and one trematode egg were seen. There were no significant differences in the prevalence of parasites between males and females and between age groups: however, infants and juveniles appeared to have a lower prevalence of Anoplocephala gorillae, and the silverbacked males appeared to have a higher prevalence of Probstmayria sp. Parasite prevalence was consistent among the five social groups studied except Susa group had a significantly lower prevalence of Anoplocephala gorillae. Trichuris trichiura, Strongyloides sp., Chilomastix sp., and Endolimax nana were identified for the first time in this population, and it is possible that these parasites were of human origin. Although there were no obvious clinical effects due to the presence of these parasites, six parasites identified (Trichuris trichiura, Strongyloides sp., Oesphagostomum sp., Trichostrongylus sp., Entamoeba histolytica, and Giardia sp.) could potentially be pathogenic. Some of the parasite products and cultured larvae could not be speciated.     

Proportions of various endocrine cells in the pancreatic islets of wood mice (Apodemus speciosus)

Using wood mice (Apodemus speciosus) captured in the wild in Niigata, we analysed the proportion of various endocrine cells in pancreatic islets for both immunohistochemical and microscopic characteristics. In both the dorsal and ventral portions of the pancreas, the centre of the pancreatic islets was occupied predominantly by insulin-positive (B) cells, surrounded by glucagon-positive (A), somatostatin-positive (D), and pancreatic-polypeptide-positive (PP) cells. Although the proportions of the various endocrine cells in pancreatic islets varied from one mouse to the next, in most animals B cells accounted for more than half of all endocrine cells. Dorsal and ventral portions of the pancreas differed in the proportions of various endocrine cells, specifically, in the A-to-PP cell ratio: the proportion of PP cells higher in the ventral portion. The same tendency is seen in humans, rats and mice. Microscopic examinations revealed morphologically distinct secretory granules in A, B and D cells. The morphology of these granules was similar that of secretory granules found in rats and mice.     

Gastritis and intestinal metaplasia in Syrian hamsters infected with Helicobacter aurati and two other microaerobes

Chronic gastritis and intestinal metaplasia associated with naturally occurring colonization by Helicobacter aurati and two other microaerobic species were observed in Syrian hamsters. Thirty-five hamsters, between 7 and 12 months of age, were evaluated from two research and three commercial facilities. Microaerobic bacteria were cultured from the hamster stomachs. These bacteria included H. aurati, a fusiform, urease-positive species; a second novel helical, urease-negative Helicobacter sp.; as well as a smaller, urease-negative Campylobacter sp. Southern blot analysis detected Helicobacter spp. DNA in the gastric tissues of all 35 hamsters; 15 hamsters also had Campylobacter sp. DNA in their gastric tissues. When examined by light microscopy, argyrophilic bacteria consistent with H. aurati or the second Helicobacter sp. were present in antral sections of 12 out of the 15 hamsters where bacteria were seen, while 9 out of the 15 hamsters had bacteria resembling the Campylobacter sp. The presence of Helicobacter spp. but not the presence of Campylobacter sp. was significantly correlated to gastritis severity (P < 0.0001 for Helicobacter spp., P = 0.6025 for Campylobacter sp.) and intestinal metaplasia, as measured by numbers of goblet cells (P = 0.0239 for Helicobacter spp., P = 0.5525 for Campylobacter sp.). Severely affected hamsters also had Giardia sp. within their metaplastic gastric pits. Hamsters with naturally occurring helicobacter-associated gastritis provide a model for studying the development of intestinal metaplasia and gastric giardiasis in H. pylori-infected humans.     

The rapid identification of Escherichia coli using Bactident E. coli

165 (93.7%) of 175 E. coli strains isolated from various materials of different animal species were correctly identified as E. coli with help of commercially available Bactident E. coli (E. Merck, D-6100 Darmstadt). Further 52 strains of the family of Enterobacteriaceae (2x Providencia sp., 2x Salmonella sp., 7 Citrobacter sp., 9 Proteus sp., 15 Klebsiella sp., 17 Enterobacter sp.) were correctly not identified as E. coli by this test. Bactident E. coli is a suitable test for rapid identification of E. coli in veterinary bacteriology.     

The rapid identification of E. coli by the detection of 3 enzymes

232 isolates of gram-negative, oxidase-negative bacteria, isolated from various samples of different animal species, were tested with help of RAPIDEC coli for the production of beta-glucuronidase, beta-galactosidase and indole. The test correctly identified 164 of 175 E. coli strains (sensitivity 93.7%) and correctly indicated that 57 of 57 isolates of the family Enterobacteriaceae (7 Citrobacter sp., 18 Enterobacter sp., 16 Klebsiella sp., 10 Proteus sp., 2 Providencia sp. and 4 Salmonella sp.) were not E. coli (specificity 100%).     

Immunohistochemical Study of Peptide-containing Nerves in the Gastrointestinal Tract of the Japanese Field Vole, Microtus montebelli

The distribution and relative frequency of nerves containing immunoreactivity for substance P (SP), vasoactive intestinal polypeptide (VIP), gastrin-releasing polypeptide (GRP), calcitonin gene-related peptide (CGRP), neuropeptide Y (NPY) and menthionine-enkephalin (MENK) were studied by immunohistochemistry in the gastrointestinal tract of the herbivorous Japanese field vole, Microtus montebelli.     

A possible role for Clostridium difficile in the etiology of calf enteritis

Clostridium difficile was investigated as a possible cause of enteritis in calves. The organism and its toxins (TcdA and TcdB), respectively, were found in 25.3% and 22.9% of stool samples from diarrheic calves. Culture positive samples were more likely than culture negative samples to be toxin positive. However, toxin positive stools were more common among nondiarrheic calves, but diarrheic calves were nearly twice as likely to be culture positive. Ribotype 078 was dominant among isolates. Salmonella sp. was isolated from both diarrheic and nondiarrheic calves, but large numbers of E. coli were found more commonly in diarrheic calves than in nondiarrheic animals. Prevalence rates for coronavirus and Cryptosporidium sp. were substantially higher in nondiarrheic calves than in diarrheic, but rates of detection of rotavirus and Giardia sp. were more nearly equal between groups. Lesions in naturally infected calves included superficial mucosal erosion with associated fibrinous exudates. Neutrophils and eosinophils infiltrated lamina propria. Large Gram-positive rods morphologically compatible with C. difficile were abundant in the colonic lumen and the organism was isolated by bacteriologic culture. Toxins were found throughout the colon. Purified toxins A and B (individually and conjointly) caused comparable lesions, as well as fluid accumulation, in ligated intestinal loops. Our findings are in substantial agreement with those of others [Rodriguez-Palacios, A., Stampfli, H.R., Duffield, T., Peregrine, A.S., Trotz-Williams, L.A., Arroyo, L.G., Brazier, J.S., Weese, J.S., 2006. Clostridium difficile PCR ribotypes in calves, Canada. Emerg. Infect. Dis. 12, 1730-1736; Porter, M.C., Reggiardo, C., Bueschel, D.M., Keel, M.K., Songer, J.G., 2002. Association of Clostridium difficile with bovine neonatal diarrhea. Proc. 45th Ann. Mtg. Amer. Assoc. Vet. Lab. Diagn., St. Louis, MO, U.S.A.] and add strength to a working hypothesis that C. difficile infection and the accompanying intoxication can manifest as diarrhea in calves. It seems clear that calves serve as multiplying hosts for this organism.     

Parasites from farmed ostriches (Struthio camelus) and rheas (Rhea americana) in Europe

During a 4-year-period, more than 500 ostriches and several rheas, all born in European countries and raised in Spain and Portugal, have been analyzed for the presence of ectoparasites and endoparasites. A total of 29 parasite species have been found, most of them of the gastrointestinal tract. Some of the helminth species found may represent spureous parasitosis, as only the eggs (of an ascarid and a trematode) were found in some samples. From the organisms identified, the ectoparasites (lice-Struthiolipeurus rheae, S. nandu; mites-Dermoglyphus pachycnemis, Gabucinia bicaudata), helminths (cestoda-Houttuynia struthionis- and nematoda-Libyostrongylus sp., Codiostomum struthionis-) and the ciliate Balantidium struthionis are known as ratite specific parasites. Capillaria eggs and larvae were also found; there are no previous records of this parasite from ostriches, and the data available do not allow to do a temptative specific diagnosis. Among protozoa, most of the species now found are described for the first time in ratites. They include organisms also found in other birds (Trichomonas gallinae, Tetratrichomonas gallinarum, Chilomastix gallinarum, Spironucleus meleagridis and Pleuromonas jaculans), and organisms whose specific status cannot be established until further analysis are performed (Cryptosporidium sp., Eimeria sp. and/or Isospora sp., Entamoeba sp. of the one-nucleate and of the eight-nucleate mature cyst groups, Endolimax sp., Iodamoeba sp., Monocercomonas sp., Retortamonas sp., Giardia sp., Blastocystis sp. and euglenids).     

The prevalence of blood parasites in helmeted guineafowls, Numida meleagris, in the Kruger National Park

Bloodsmears were taken from separate groups of five helmeted guineafowls, Numida meleagris, shot at approximately monthly intervals at Skukuza and near Lower Sabie in the Kruger National Park during the period August 1988 to August 1990. Ninety-eight (86%) of 114 guineafowls had single or multiple infections of Aegyptianella sp., Haemoproteus pratasi, Hepatozoon sp., Leucocytozoon neavei, Plasmodium circumflexum and Trypanosoma numidae. The apparent seasonal prevalence of Aegyptianella sp., H. pratasi and L. neavei, the three most commonly occurring parasites (42%, 49% and 56% of birds infected respectively), is probably dependent on the presence of their respective vectors.     

Multidrug and multispecies resistance in sheep flocks from São Paulo state, Brazil

The economic importance of sheep production is increasing worldwide simultaneously with the emergence of parasitic resistance. This study aimed to survey the current situation of management practices and parasite resistance in sheep flocks in S?o Paulo state, Brazil. A questionnaire was given to 35 sheep farmers to obtain information related to flock management practices. Of these flocks, 30 were submitted to the fecal egg count reduction test (FECRT) with at least one of the five following anthelmintics: albendazole, closantel, ivermectin, levamisole, and moxidectin, for comparison against an untreated control group. In the survey, the median number animals per flock was 301, mainly of the Santa Ines breed (in 75.8% of the flocks) and crossbred animals (in 54.5% of the flocks). The predominant farming system was semi-intensive (82.9%), using rotational grazing (80%). Selective treatment was based on FAMACHA grade (47.1%) and in clinical signs (41.2%). The most often applied anthelmintics were macrocyclic lactones (42.9-54.2% in the last three applications). Considering the anthelmintics employed in this study, 10.7% of the farms' flocks were resistant to three, 35.7% to four, and 53.6% to all five anthelmintics. The main helminth genera observed before and after treatments were Haemonchus sp. (75.8%) and Trichostrongylus sp. (19.1%), but all observed genera (Cooperia sp., Oesophagostomum sp., and Strongyloides sp.) were detected by the FECRT. Considering efficacy values less than or equal to 90% in the FECRT as resistant, 100% of flocks were resistant to albendazole and ivermectin, 96.6% to moxidectin, 92.9% to closantel, and 53.6% to levamisole. It is thus possible to conclude that multidrug resistance is widespread in sheep flocks in S?o Paulo state, Brazil, and this involves all prevalent helminth genera.