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1.
We previously reported that alcoholic extracts of Sophora flavescens and Torilis japonica from South Korea demonstrated good efficacy in reducing replication of Toxoplasma gondii and Neospora caninum. To characterize the chemical component associated with anti-protozoal activity, specific fractions were isolated by high performance liquid chromatography (HPLC) and used for in vitro testing. These fractions were evaluated in vitro against T. gondii and N. caninum. Fractions of the herb extracts were serially diluted to final concentrations of 2.850 to 0.356 ng/ml in medium and added to wells containing replicating T. gondii and N. caninum. To determine the ability of each fraction to inhibit parasite proliferation, 3H-uracil incorporation was used to determine parasite replication. In cultures infected with T. gondii, a fraction of T. japonica (TJ2) inhibited T. gondii proliferation by 99.2, 94.4, 88.6 and 27.0% in the range from 2.850 to 0.356 ng/ml. Four fractions of S. flavescens (SF1-SF4) inhibited T. gondii proliferation by 99.6-60.6, 96.9-48.1, 92.3-68.2 and 95.4-52.9% in the range from 2.850 to 0.356 ng/ml. In cultures infected with N. caninum, a fraction of T. japonica (TJ2) inhibited N. caninum proliferation by 98.3, 95.5, 79.7 and 30.6% in the range from 2.850 to 0.356 ng/ml. Four fractions of S. flavescens (SF1-SF4) inhibited N. caninum proliferation by 97.1-25.9, 94.8-35.5, 95.9-33.7 and 95.4-49.4% in the range from 2.850 to 0.356 ng/ml. These fractions of T. japonica and S. flavescens extracts are currently undergoing in vivo evaluation in experimentally infected mice.  相似文献   

2.
Ionophorous antibiotics have been popularly used in the treatment of avian coccidiosis. Tissue residue of these antibiotics may be found in poultry, we have sought safe alternative anticoccidial herbal materials for the control of avian coccidiosis. Efficacy of extracts from 15 different herbs, including Bupleurum chinese DC, Sophora flavescens Aiton, and Artemisia annua Linne was tested against Eimeria tenella. One-day-old broiler chicks were infected with a USDA reference book of E. tenella, and administered various herbal extracts.Survival rates, lesion scores, body weight gains, bloody diarrhea, and oocysts excretions were investigated at the first and the second week after infection. Bloody diarrhea in the S. flavescens and Sinomenium acutum treated groups was milder than that of the other infected groups. Survival rates in the groups treated with Ulmus macrocarpa (100%), Pulsatilla koreana, Torilis japonica, Artemisia asiatica and S. flavescens (90%) were higher than that of the infected control group (70%). Lesion scores in the groups treated with U. macrocarpa (1.40+/-1.14) or Pulsatilla koreana (1.60+/-1.82) were significantly lower than those of the infected control group (3.00+/-1.10). During the first week after infection, the weight gains in the groups treated with Quisqualis indica (232.9+/-43.5 g), S. flavescens (214.4+/-46.1 g) and S. acutum (211.3+/-29.4 g) were significantly higher than the infected control group (172.4+/-17.6 g).In conclusion, the data of the survival rates, bloody diarrhea symptoms, lesion scores, body weight gains and oocyst excretions indicate that the extract of S. flavescens was the most effective. P. koreana, S. acutum, U. macrocarpa and Q. indica were also effective. Further research on the above herbal materials will be carried out by the authors by chemical analysis of the extracts.  相似文献   

3.
Sera from Cervidae were tested for the presence of antibodies against Neospora caninum using ELISA; and against Toxoplasma gondii using SAG1-ELISA and a commercially available agglutination test. The T. gondii seroprevalence was 52% (38/73) in roe deer (Capreolus capreolus), 0% in bred fallow deer (0/4) (Dama dama) and red deer (0/7) (Cervus elaphus). We found 2.7% of the roe deer samples and none of the bred deer samples positive for N. caninum. Brain samples from wild roe deer, red deer and red foxes (Vulpes vulpes) were tested for the presence of T. gondii and N. caninum DNA using multiplex real-time PCR. We detected T. gondii in 18.8% (57/304) of the red foxes and in 1 of the 33 deer samples. N. caninum was found in 6.6% of the red foxes and in 2 roe deer samples. Twenty-six of the T. gondii positive DNA extracts from the red fox samples were genotyped. Twenty-five were type II and only one was found to be type III.  相似文献   

4.
Immunohistochemical diagnosis of Neospora caninum in tissue sections   总被引：10，自引：0，他引：10  
An avidin-biotin-peroxidase complex immunoperoxidase staining method was developed to detect Neospora caninum in formalin-fixed, paraffin-embedded tissue sections. Specific antiserum to N caninum was made in rabbits and used to probe tissues from dogs naturally and experimentally infected with N caninum. The test detected tachyzoites and bradyzoites of N caninum. A reaction was not observed to Toxoplasma gondii, Hammondia hammondi, Sarcocystis cruzi, S capricanis, S tenella, Besnoitia jellisoni, Caryospora bigenetica, Hepatazoon canis, Atoxoplasma sp, or the organism causing canine dermal coccidiosis. When antiserum made in rabbits to T gondii was used in the test, reaction to N caninum was not observed.  相似文献   

5.
OBJECTIVE: To determine serologic prevalence of Sarcocystis neurona, Toxoplasma gondii, and Neospora caninum in horses in Brazil. DESIGN: Prevalence survey. ANIMALS: 101 Thoroughbreds in Brazil. PROCEDURE: Blood samples were obtained from horses and tested for serum antibodies against S neurona by use of an immunoblot procedure with culture-derived S neurona merozoites as antigen, and for serum antibodies against T gondii and N caninum by use of a modified agglutination test with formalin-preserved tachyzoites and mercaptoethanol. RESULTS: Antibodies against S neurona and T gondii were detected in 36 and 16 of 101 horses, respectively. Cross-reactivity between antibodies against T gondii and S neurona was not detected. Antibodies against N caninum were not detected in any samples. CONCLUSIONS AND CLINICAL RELEVANCE: The high prevalence of antibodies against S neurona detected in clinically normal horses emphasizes the importance of examining CSF for antibodies when establishing a diagnosis of equine protozoal myeloencephalitis.  相似文献   

6.
Llamas (Lama glama) are South American camelids described as intermediate hosts of Neospora caninum, Toxoplasma gondii and Sarcocystis aucheniae. Due to the potential role of these protozoan infections as a cause of economic losses, the aim of this study was to determine the seroprevalence for T. gondii, N. caninum and Sarcocystis sp. in llamas from Argentina. Serum samples from 308 llamas (>2 years old) were collected between 2005 and 2007. A total of 55 farms located in six departments of Jujuy province, Argentina were sampled. Presence of antibodies to N. caninum, T. gondii and Sarcocystis sp. was determined by the indirect fluorescent antibody test (IFAT). For Sarcocystis, 2 different bradyzoites-based antigens were prepared using S. aucheniae and S. cruzi. Sera were tested at dilutions 1:25 and 1:50. Antibodies to N. caninum were found in 4.6% serum samples. Fifty percent of departments and 14.5% of farms had positive animals. Antibodies to T. gondii were found in 30% of samples, distributed in 66% of departments and 43.6% of farms. Antibodies to Sarcocystis sp. were detected in 96% of samples and all departments and farms had positive animals, suggesting frequent contact between llamas and canids. Co-infection with N. caninum, T. gondii and Sarcocystis sp. was also recorded. Low seroprevalence of N. caninum in llamas detected in this study could be related to climatic and geographical conditions that limit cattle breeding activity, reducing the source of infection for definitive hosts. Seroprevalence of T. gondii and the positive animal distribution suggest frequent contamination of grass with felid faeces. In conclusion, this is the first report of combined seroprevalence for N. caninum, T. gondii and Sarcocystis sp. in llamas. Further studies are needed to determine the potential role of these protozoan infections as cause of abortion in Argentina as well as presence of these protozoans in llama meat used for human consumption.  相似文献   

7.
Neosporosis and toxoplasmosis are two important infections in young and adult sheep, leading to low production and abortion. This study aimed to determine the frequency of antibodies to Toxoplasma gondii and Neospora caninum in sheep from the eastern region of S?o Paulo State, Brazil. Serum samples (382) were collected from the sheep and assayed for T. gondii through modified agglutination test (MAT) and indirect fluorescent antibody test (IFAT), and for N. caninum antibodies, through IFAT, with cut-off titers equal to 16 (T. gondii) and 25 (N. caninum). All frozen samples were sent to the Center for Zoonoses Research (NUPEZO), Department of Veterinary Hygiene and Public Health (DHSVP), FMVZ, UNESP, for serological tests. A total of 71/382 (18.6%) samples reacted to T. gondii, especially at titers 16 (28; 39.4%), 64 (15; 21.1%), 256 (21; 29.6%) and 1024 (6; 8.5%) by MAT, and 16 (34; 47.9%), 64 (18; 25.4%), 256 (14; 19.7%) and 1024 (5; 7%) by IFAT. As regards N. caninum, 49/382 (12.8%) samples reacted at titers 25 (17; 34.7%), 50 (11; 22.5%), 100 (11; 22.5%), and ≥ 200 (10; 20.4%). These animals presented infection but no clinical signs. Six and ten animals had high titers for toxoplasmosis and neosporosis. No significant association was observed between antibodies for both parasites (P=0.535) according to Fisher's exact test, and no correlation was found between T. gondii (MAT) and N. caninum antibody titers (r=-0.0068; P=0.895), T. gondii (IFAT) and N. caninum antibody titers (r=-0.0025; P=0.961). Thus, T. gondii and N. caninum infections were observed in farms located in S?o Paulo State, where sheep play an important economical role for the national and regional business.  相似文献   

8.
Neospora caninum and Toxoplasma gondii are cyst-forming coccidian parasites of human and veterinary clinical relevance. In vitro cultivation of the protozoans using Vero cells is usually performed in order to produce antigenic materials. Quantitative and qualitative comparisons of Vero cells grown in RPMI medium supplemented either with foetal calf serum (FCS), horse serum (HS) or a specific serum-free additive (DefCell) were performed. A serum-free cell culture system used to propagate N. caninum (NC-1 isolate) and T. gondii tachyzoites (Rh stain) were compared with the other two cell culture systems. FCS supplemented media was found to be more effective than the others in promoting Vero cells and N. caninum tachyzoites. However, it was found unable to support adequate T. gondii tachyzoite proliferation. Vero cells, T. gondii and N. caninum tachyzoite production gave similar growth patterns with either HS or DefCell supplemented media. Defcell was considered as a good alternative to supplement culture medium.  相似文献   

9.
Liu J  Cai JZ  Zhang W  Liu Q  Chen D  Han JP  Liu QR 《Veterinary parasitology》2008,152(3-4):330-332
Neospora caninum is a protozoan parasite and is closely related to Toxoplasma gondii, but they are antigenically different. N. caninum and T. gondii infection in a variety of animals such as cattle, dogs, and cats has been reported, but there is little information on the infection of these parasites in domestic yaks. Seroprevalence of antibodies to T. gondii and N. caninum in yaks (Bos grunniens) from eight regions of Qinghai, China were investigated by the indirect agglutination test (IAT) and ELISA, respectively. A total of 112 (11.8%) of 946 serum samples were positive for antibodies to T. gondii, and 21 samples (2.2%) were positive to N. caninum. Two of the yaks had antibodies to both parasites. There was no apparent association of T. gondii infection with age of the animals. The results indicate that T. gondii infection is prevalent in Chinese yaks in most parts of Qinghai province and N. caninum infection rate in the same species is relatively low. This is the first large study showing the infection of T. gondii and N. caninum in domestic yaks.  相似文献   

10.
Little is known about the epidemiology of Neospora caninum in wild mammal populations. It has been suggested that a sylvatic cycle exists for N. caninum. Dogs and potentially other canids are a definitive host for N. caninum. The present study was done to determine the prevalence of antibodies to N. caninum in a population of gray foxes (Urocyon cinereoargenteus) from a nonagricultural setting in South Carolina. We also determined the prevalence of antibodies to Toxoplasma gondii in these animals. Antibody levels were measured in direct agglutination tests using either N. caninum or T. gondii formalin-fixed tachyzoites as antigen. Four (15.4%) of the 26 gray foxes had titers to N. caninum. Titers to N. caninum were low being 1:25 in three gray foxes and 1:50 in the fourth gray fox. Antibodies to T. gondii were observed in 16 (61.5%) gray foxes. Titers to T. gondii were usually >1:50 and two gray foxes had titers of 1:1600. Results of this study indicate that gray foxes have more exposure to T. gondii than to N. caninum in this environment.  相似文献   

11.
Sera from 68 nondomestic captive and free-ranging felids from southern Africa were tested for antibodies to Neospora caninum and Toxoplasma gondii by the indirect fluorescent antibody test. Four of the 68 (5.9%) serum samples were positive for antibodies to N. caninum, with titers ranging from 1:50 to 1:200. All other animals were negative for antibodies to N. caninum at a dilution of 1:50. Fifty of the 68 (74%) serum samples tested positive for antibodies to T. gondii, with titers ranging from 1:50 to 1:26,500. Four animals tested positive for antibodies to both N. caninum and T. gondii. None of these animals displayed clinical signs of disease. Results of this study indicate that nondomestic felids in southern Africa have been exposed to, and are likely infected with, N. caninum and T. gondii.  相似文献   

12.
A serological survey for antibodies against Neospora caninum in cattle, goats and farm dogs in Taiwan was carried out. Sera of 613 cattle from 25 dairy farms, 24 goats from six goat farms and 13 dogs from six dairy cattle farms were tested for antibodies against N. caninum using indirect fluorescence antibody test (IFAT). The same sera were also tested for antibodies against Toxoplasma gondii using latex agglutination test. Of the 613 cattle sera, 44.9% (275/613) were found to have antibodies against N. caninum. Among these 275 positive cattle, 77 also possessed antibodies against T. gondii. Nevertheless, 92 cattle which were negative for N. caninum showed antibodies against T. gondii. Of the 24 goat sera tested, none was found to be positive for N. caninum but 50% (12/24) were positive for T. gondii. Of the 13 farm dogs tested, three were found to possess antibodies against N. caninum, two of which tested negative for T. gondii antibodies. Besides sera, antibodies to N. caninum in cattle could be observed in the milk, vaginal secretion and saliva. However, the order of higher frequency of antibodies detection is in sera, milk, vaginal secretion and saliva. This is the first demonstration of the presence of antibodies to N. caninum in vaginal secretion and saliva of cattle. A 50microm cyst was observed in the brain of one of the 13 prednisolone-treated SPF ICR mice which had been peritoneally inoculated 4 months earlier with the brain homogenate of a serologically N. caninum positive but T. gondii negative cattle. Thus, we have confirmed for the first time the presence of N. caninum in Taiwan and also observed that it is widespread among dairy cattle and farm dogs.  相似文献   

13.
The main purpose of the present study was to investigate the occurrence of antibodies against T. gondii and N. caninum in captive maned wolves from Brazil, considering that little information is available at the literature about infections by these parasites in this wild animal. Serum samples were obtained from 59 maned wolves originated from six zoos and from one ecological reserve of the southeastern and midwestern regions of Brazil. To detect IgG antibodies against T. gondii, an ELISA protocol was used and the results were expressed as ELISA reactivity indexes (EI). Serology for N. caninum was carried out by indirect fluorescent antibody test (IFAT) and cut-off titers were established at 1:25 dilution. From the total of the analyzed samples, 44 (74.6%) were seropositive for T. gondii and only 5 (8.5%) for N. caninum. Seropositivity for T. gondii ranged from 0 to 100% in the seven different origin locals, with rates over 50% among the six zoos, whereas no positivity was found in the samples from ecological reserve. For N. caninum, seroprevalence varied from 0 to 50% in the different locals, with the highest rates also detected in zoos. Seroprevalence for T. gondii was strongly related with age, with rates significantly higher among adult wolves (91.7%) when compared to newborn or young animals. Seropositive samples for N. caninum were found predominantly in adult wolves. For both parasites, seroprevalence did not show a significant distinction in relation to gender. Although seroprevalence for T. gondii was significantly higher when compared to N. caninum in the Brazilian captive maned wolves tested, these findings reflect the great exposure of this species to T. gondii and, in lower extension, to N. caninum. Also, the present study demonstrated for the first time the presence of antibodies to N. caninum in wild life from South America.  相似文献   

14.
Little information is available about the seroprevalence of Toxoplasma gondii and Neospora caninum infections in goats in Romania and even in Europe. During 2007-2010, 735 serum samples were collected from dairy goats located in 4 historical regions (Cri?ana, Maramure?, Transylvania and Muntenia) of Romania. Sera were analyzed for T. gondii and N. caninum antibodies (IgG type) by enzyme-linked immunosorbent assay (ELISA) using two commercial kits (Chekit Toxotest Antibody ELISA and Chekit Neospora caninum Antibody ELISA; Idexx-Bommeli, Switzerland). Three hundred and eighty-eight out of 735 (52.8%) goats presented T. gondii antibodies and 12 out of 512 (2.3%) goats had N. caninum antibodies. The high seroprevalence of T. gondii suggests that infection with this parasite is common in dairy goats in Romania, and less common the infection with N. caninum. This is the first time that infection with N. caninum in goats has been reported in Romania and the first extended study on seroepidemiology of T. gondii.  相似文献   

15.
In September 2004, a neonatal sea otter pup was found alive on the beach in northern Monterey Bay, CA. Efforts to locate the mother were unsuccessful. Due to a poor prognosis for successful rehabilitation, the pup was euthanized. Postmortem examination revealed emaciation, systemic lymphadenopathy and a malformation of the left cerebral temporal lobe. On histopathology, free tachyzoites and tissue cysts compatible with Toxoplasma gondii were observed in the brain, heart, thymus, liver, lymph nodes and peri-umbilical adipose. The presence of T. gondii within host tissues was associated with lymphoplasmacytic inflammation and tissue necrosis. Immunofluorescent antibody tests using postmortem serum were positive for anti-T. gondii IgM and IgG (at 1:320 and 1:1280 serum dilution, respectively), but were negative for IgG directed against Sarcocystis neurona and Neospora caninum (<1:40 each). Brain immunohistochemistry revealed positive staining for tachyzoites and tissue cysts using antiserum raised to T. gondii, but not S. neurona or N. caninum. T. gondii parasite DNA was obtained from extracts of brain and muscle by PCR amplification using the diagnostic B1 locus. Restriction enzyme digestion followed by gel electrophoresis and DNA sequencing confirmed the presence of Type X T. gondii, the strain identified in the majority of southern sea otter infections.  相似文献   

16.
Serum samples from 597 sheep from S?o Paulo State, in the southeastern region of Brazil, were tested to determine the prevalence of antibodies directed against Toxoplasma gondii (> or = 1:64) and Neospora caninum (> or = 1:50) using the indirect fluorescent antibody test (IFAT). The animals were divided into three groups based on their age: < or = 1 year, 1-4 years, and > or = 4 years. Antibodies to T. gondii were observed in 34.7% of the samples with titers ranging from 64 to 16,384 and IgG antibodies directed against N. caninum were observed in 9.2%, with titers ranging from 50 to 3200. Only 3.5% of the sheep were positive for both agents. All farms had at least one positive animal for T. gondii, and 26 of the 30 farms had at least one positive animal for N. caninum. An association between seroprevalence and age was observed for T. gondii (P = 0.001), but not to N. caninum (P = 0.343). It was not possible to associate seroprevalence to T. gondii and the presence of domestic or feral cats, since in all farms there was at least one positive sheep. There was no association between seropositivity to N. caninum and the presence of domestic (P = 1.000) and feral dogs (P = 0.550).  相似文献   

17.
Toxoplasmosis and neosporosis have been recognized as economically important diseases with considerable impact on the livestock industry. Considering the scarce information on the occurrence of Toxoplasma gondii and Neospora caninum infections in sheep from Uberlandia, Minas Gerais State, Brazil, this study aimed to investigate the frequency of antibodies against these parasites in sheep sera from this region by using different serological methods. A total of 155 sheep serum samples were analyzed by the indirect fluorescence antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) for the detection of IgG against T. gondii and N. caninum. Seroreactivity by IFAT showed 80% of samples with titers between 512 and 2048 for T. gondii (cutoff ≥ 64) and 78% presenting titers between 50 and 200 for N. caninum (cutoff ≥ 50). Seroreactivity by ELISA showed 75% of samples with ELISA index (EI) between 2.0 and 3.0 for T. gondii (cutoff ≥ 1.3) and 54% presenting EI between 1.3 and 2.0 for N. caninum (cut off ≥ 1.3). Discordant results by both tests were analyzed by immunoblot, resulting in a total seropositivity of 61% for T. gondii and 23% for N. caninum, with 41% to T. gondii only, 3% to N. caninum only, and 20% to both parasites. There was a significant positive association between seropositivity to T. gondii and age over one year (P<0.001), but such association was not found for N. caninum infection. In conclusion, as T. gondii and N. caninum infections are simultaneously present in sheep flocks of this region, it should be emphasized the importance to carry out a regular monitoring of Toxoplasma infection due to its high prevalence, its zoonotic potential and induction of reproductive disorders leading to economic losses. For neosporosis, sheep farmers should be instructed about the presence of the parasite in the flock, its risk factors and potential abortifacient role in sheep. Differential flock management could be valuable tool to establish the association of serological positivity and reproductive disease induced by N. caninum in sheep.  相似文献   

18.
The enzyme-linked immunosorbent assay (ELISA) was used to examine cross-reactivity of Neospora caninum with Toxoplasma gondii and Hammondia heydorni. Anti-T. gondii mouse and cat sera cross-reacted with N. caninum soluble antigen (NLA), but not with the recombinant surface antigen (NcSRS2). Anti-H. heydorni dog sera showed no cross-reactivity with either the NLA antigen or the NcSRS2. Lack of cross-reactivity between anti-H. heydorni sera and N. caninum antigens, and the cross-reactivity of anti-T. gondii sera with the NLA suggest that N. caninum has common antigens to T. gondii except for NcSRS2 based on serology. In light of several studies suggesting a closer relationship between N. caninum and H. heydorni than with T gondii, examination of serological cross-reactivity with N. caninum may be necessary to further classify the parasites in addition to molecular and morphological studies and clarification of the life cycle.  相似文献   

19.
Neospora caninum and Toxoplasma gondii are closely related cyst-forming apicomplexan parasites identified as important causes of reproductive failure in cattle and small ruminants, respectively. Protozoan abortion in small ruminants is traditionally associated with T. gondii, but the importance of N. caninum remains uncertain. The aim of this study was to investigate the presence of N. caninum and T. gondii infections in abortion cases in small ruminants submitted for diagnosis. For this purpose, 74 ovine and 26 caprine aborted foetuses were recovered from different areas in Spain. Foetal histopathology was used to detect the presence of protozoal-associated lesions in brain. The presence of N. caninum and T. gondii was confirmed by PCR. Protozoal infection was detected in 17 out of 100 (17%) foetuses examined by at least one of the diagnostic techniques used. Lesions suggestive of protozoal infection were observed in 10.8% (8/74) and 15.4% (4/26) of the ovine and caprine abortions respectively. N. caninum and T. gondii infection was detected by PCR in 6.8% (5/74) and 5.4% (4/74) of sheep foetuses, respectively, of which five showed protozoal-associated lesions. N. caninum DNA was detected in 11.5% (3/26) of goat foetuses, of which two showed protozoal-associated lesions, whereas T. gondii DNA was detected in one goat foetus with no lesions. The simultaneous presence of N. caninum and T. gondii DNA was detected in one sheep foetus with severe lesions. This study demonstrates that N. caninum plays a significant role in abortion in small ruminants in the studied population. In addition, our results highlight the importance of differentiating between protozoa whenever characteristic lesions are observed.  相似文献   

20.
A seroepidemiological survey of Neospora caninum and Toxoplasma gondii in cattle and water buffaloes was carried out in the People's Republic of China. Serum samples were obtained from dairy (n=262, 9 herds in 9 provinces) and beef cattle (n=10, 1 herd) and water buffaloes (n=40) in China. All sera were tested for antibodies to N. caninum and T. gondii by an enzyme-linked immunosorbent assay (ELISA) and an indirect agglutination test (IAT), respectively. The overall seroprevalence of N. caninum in dairy cattle was 17.2% (45/262), and the herds seroprevalence of N. caninum was 88.9% (8/9), and antibodies to T. gondii were present in 6 cows (2.3%). None of the cows had antibodies against both T. gondii and N. caninum. Antibodies to T. gondii or N. caninum were not found in beef cattle or water buffaloes. The seroprevalence of N. caninum in aborting cows (20.2%) was higher than that in non-aborting cows (16.6%) with an odds ratio of 1.26 (95% CI, 0.54-2.95), but the difference was not statistically significant (P>0.05). There was no apparent association of N. caninum seropositivity with age or number of pregnancies. This is the first report on the seroprevalence of N. caninum in cattle and water buffaloes in China.  相似文献   

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