Control ofTheileria sergenti infection by vaccination

Bovine piroplasmosis caused byTheileria sergenti is a major cause of economic loss in grazing cattle in Japan. Infected calves show chronic anaemia with intraerythrocytic piroplasms and occasionally die in severe cases. We found that parasite stocks and isolates consist of genetically and antigenically mixed populations. To differentiate parasite populations bearing 3 allelic forms of p32/34, an immunodominant piroplasm surface protein, 3 sets of oligonucleotide primers were designed to amplify either of 3 alleles by polymerase chain reaction (PCR). By using this allele-specific PCR, we found that the majority ofT. sergenti-infected calves in Japan harbored mixed parasite populations bearing C and I type parasites. To controlTheileria infection, we produced 2 vaccine candidates: recombinant baculovirus p32 and synthetic peptide containing Lys-Glu-Lys (KEK) motif. Immunization with either recombinant p32 or synthetic peptide containing KEK sequences with Freund’s complete adjuvant resulted in low parasitemia and reduced the clinical symptoms compared to control calves. Interestingly, the parasite with the p32 allelic form corresponding to the one used as the immunogen was suppressed.

     

Comparisons of the resistance to Japanese theileriosis among Santa Gertrudis × Japanese Shorthorn F1, Japanese Shorthorn and their reciprocal crosses

To improve the resistance of Japanese Shorthorn (JS) calves to the hemoprotozoan parasite Theileria sergenti (Ts), Santa Gertrudis (SG) × JS F₁ and its reciprocal backcross (F₁ × JS and JS × F₁) calves were produced. All the calves were born from February to April and kept on pasture with their dams from May to October. Blood samples were collected biweekly until 18 weeks of grazing and two indices of resistance to the parasite, percentage of parasitized erythrocytes (PE) and hematocrit value (Ht) were monitored. There was a weak negative correlation between daily gain (DG) on pasture and the average PE (P < 0.1), and was a significantly positive correlation between DG on pasture and the average Ht (P < 0.01) in the experimental period. Although maximum PE observed at six or eight weeks of grazing period was not significantly different among the four mating types, SG × JS F₁ calves exhibited a significantly larger decrease in PE compared to purebred JS after the peak PE. F₁ calves also showed maximum Ht through the grazing period. There were no apparent differences in the two indices among the two reciprocal backcrosses and purebred JS. These results indicated that Ts infection and subsequent decrease in Ht affected the DG of calves in the grassland. The present study also demonstrated that the introduction of the SG gene into JS was effective at the F₁ level in regard to the resistance to theileriosis, and that in the backcrosses an improvement in resistance could not be expected.     

Susceptibility of Chinese hamsters (Cricetulus griseus) to the infection of Babesia microti

Chinese hamsters were examined for the susceptibility to the infection with Babesia microti based on the hematological parameters during the course of infection. A marked decrease in the RBC count, Ht value, Hb concentration, and an increase in WBC count due to the development of neutrophils or monocytes were recognized with the progress of parasitemia. Remarkable clinical findings were anemia and persistent infection with a low level of parasite burden in the chronic and convalescent stages. From these findings, it was concluded that Chinese hamsters were susceptible to infection with B. microti and would be useful for infection examination with the parasite.     

Effect of total alkaloid ofPeganum harmala L. in the treatment of experimental haemosporidian infections in cattle

Cattle experimentally infected withBabesia bigemina orTheileria sergenti or mixed infestations of the two parasites were treated with Total Alkaloid ofPeganum harmala L. The results showed that treatment was effective againstB. bigemina infection, had a marked effect on the course of infection withT. sergenti and some effect on the course of the mixed infection.

     

High susceptibility of Djungarian hamsters (Phodopus sungorus) to the infection with Babesia microti supported by hemodynamics

As the comparative study was carried out on the susceptibility by the pursuit of parasitemia among the Djungarian, Syrian, and Chinese hamsters as well as BALB/c mice infected with the Syrian hamster-adapted Babesia microti strain, and Djungarian hamsters showed the highest parasitemia among them. Then, the other hematological parameters were pursued in the Djungarian hamsters infected with the hamster-adapted B. microti strain. Remarkable symptoms observed were hemoglobinuria clinically, anemia hematologically, and splenomegaly macroscopically during all over the observation period for 24 weeks post infection (PI). Parasitemia began to rise at 2 weeks and peaked at 4 weeks PI. After that, parasitemia decreased gradually but was maintained with a level of about 10% on average until 24 weeks PI at the end of the experiment. A decrease in the RBC count, Hb, and PCV, and an increase in the reticulocyte and WBC counts due to the development of immature neutrophils, lymphocytes and monocytes were recognized together with a rise of parasitemia. The hamsters had macrocytic hypochromic anemia due to the increase of MCV and the decrease of MCHC in the growth phase of the parasite. It was considered that the Djungarian hamsters will be useful for the infection examination, isolation, maintenance, and passage of B. microti in laboratory.     

Relationship between herd size and annual prevalence of and primary antimicrobial treatments for common diseases on dairy operations in the United States

Dairy operations in states representing 86% of the United States’ national dairy herd were surveyed regarding the occurrence of common dairy diseases or disorders (digestive, respiratory, gastrointestinal, lameness, mastitis, navel infection) in specified production groups (weaned heifers, unweaned heifers, adult cows), and the most common antimicrobial preparation used to treat these conditions. Within disease and production group, disease frequency and characteristics of primary antimicrobial (drug class, availability, spectrum of activity, extra-label usage, withdrawal times) were compared among herd sizes (30–99, 100–499, and ≥500 cows) using chi-square or t-tests that accounted for the sampling design. The most common diseases were mastitis and lameness in cows, and gastrointestinal disease in unweaned calves, affecting 16, 11, and 15% of the target populations, respectively. Herd-level disease prevalence tended to increase as herd size increased, whereas within-herd prevalence tended to decrease as herd size increased. Broad-spectrum antimicrobials were selected as primary treatments by a majority of operations for nearly all diseases surveyed. When treating gastrointestinal disease, navel infection, or reproductive disorders, a majority of operations selected primary treatments not specifically labeled for that condition. Selection of over-the-counter preparations tended to decrease as herd size increased, whereas selection of broad-spectrum preparations tended to increase with herd size.     

Increased erythrophagocytic activity of macrophages in dogs with Babesia gibsoni infection

To elucidate the mechanism of anemia caused by Babesia gibsoni infection in dogs, the erythrophagocytic activity of macrophages in infected dogs was investigated in vitro. In the present study, macrophages obtained from peripheral blood (PB-macrophages) and bone marrow (BM-macrophages) of splenectomized dogs with chronic B. gibsoni infection were examined. The BM-macrophages in the splenectomized dogs with chronic babesiosis exhibited an increased erythrophagocytic activity compared with those from splenectomized, non-infected dogs. In the infected dogs, erythrophagocytic activities of macrophages against both auto- and iso-erythorcytes from normal dogs were almost the same. Administration of an anti-protozoal drug, diminazene diaceturate, resulted in a decrease of the erythrophagocytic activity of BM-macrophages associated with an increase of the hematocrit value in splenectomized dogs with chronic babesiosis. In splenectomized dogs with acute babesiosis, erythrophagocytic activity of BM-macrophages was also elevated. Such a phenomenon was not, however, observed in splenectomized dogs with onion-induced hemolytic anemia. These results suggest that the erythrophagocytic ability of macrophages in the infected dogs might be accelerated by parasites per se through an unknown mechanism, resulting in severe anemia in spite of low parasitemia.     

奶牛副结核病的诊断方法研究

副结核是一种慢性传染性细菌疾病,病原为禽分支杆菌副结核亚种,主要感染牛、羊,造成病畜小肠和其他器官损伤,引发腹泻、体重降低、营养不良、贫血甚至死亡。奶牛副结核病可导致奶牛死淘率增加、产奶量下降、饲料转化率降低、繁殖能力下降、屠宰率降低以及增加其他病原的感染率,从而对奶牛养殖业造成重大的经济损失。副结核分支杆菌主要通过粪口途径进行传播,也可通过子宫进行垂直传播。副结核分支杆菌通过在小肠黏膜下层中的巨噬细胞中持续感染来逃避免疫系统,在大多数情况下,这种隐性感染至少会持续两年,然后细菌开始脱落并出现临床症状。副结核分支杆菌感染进程缓慢,管理不当会使易感犊牛发病率更高,这使得牛副结核病的防控具有很大挑战性,尤其是在季节性产犊牛群中。本文对奶牛副结核病的概念、可造成的经济危害以及诊断方法进行阐述,旨在为奶牛副结核病的临床诊断与新型诊断方法的研制提供参考。     

中药防治奶牛乳房炎的研究进展

乳房炎是奶牛的一种常见病,是导致牛奶产量和质量下降的主要原因之一。目前,奶牛乳房炎的防治多以抗生素为主,但抗生素的滥用不仅导致了乳汁中病原菌耐药性的产生,同时也造成了乳汁中抗生素的残留。中药由于无残留、无毒副作用和较少出现耐药性等特点而备受科研学者的关注。本文主要从奶牛乳房炎的危害、中药防治奶牛乳房炎的研究现状和作用机理三个方面探讨了当前中药防治奶牛乳房炎的研究进展,旨在为临床上奶牛乳房炎的防治提供客观的参考。     

Status of oxidative stress,trace elements,sialic acid and cholinesterase activity in cattle naturally infected with Babesia bigemina

Babesia bigemina infection (also known as Texas fever) is reported as the most prevalent and main causative agent of bovine babesiosis, worldwide. The current study was undertaken to assess indicators of oxidative stress including activities of antioxidant enzymes and total antioxidant capacity (TAC), oxidation status of biomolecules and serum levels of trace elements as well as indicators of inflammation including sialic acid (SA) contents and cholinesterase activity in cattle naturally infected with B. bigemina. An infected group comprised of 20 crossbred Holstein cattle (3–4 years old) were diagnosed to be positive by both microscopy and nested PCR assay. The infected animals were subdivided into two groups according to their parasitemia rates (<20 % and>20 %). Furthermore, 10 healthy cattle were included as the control. The infection caused severe anemia in a parasitemia-burden dependent fashion. The activities of catalase and glucose-6-phosphate dehydrogenase as well as the levels of TAC, zinc, selenium, copper and manganese were significantly decreased as the parasitemia increased, accordingly. However, the activity of superoxide dismutase as well as the levels of malondialdehyde, protein carbonylation, DNA damage and iron, were significantly elevated in a parasitemeia-burden dependent manner. Additionally, glutathione peroxidase activity was significantly elevated with the lower rate of parasitemia, but the higher rate had no significant effect as compared to control. Moreover, total, protein and lipid binding SA contents were significantly increased but the activities of acetylcholinesterase and butyrylcholinesterase were significantly reduced, parasitemia dependently. Conclusively, the infection was remarkably associated with the induction of anemia, oxidative stress and inflammation.     

Comparison of the effect of T-2 toxin with that of dexamethasone or cyclophosphamide on resistance to Babesia microti infection in mice

The effect of T-2 toxin on host resistance to acute and latent Babesia microti infections was evaluated in mice and was compared with the effects of the immunosuppressive drugs dexamethasone and cyclophosphamide. Mice with acute or latent B microti infection were treated with 2 mg of T-2 toxin/kg of body weight, 0.2 mg of dexamethasone/kg, or 30 mg of cyclophosphamide/kg daily for 5 days. Treatment with dexamethasone or cyclophosphamide caused significant (P less than 0.05) increases in Babesia parasitemia during acute infection and significantly (P less than 0.05) prolonged the duration of parasitemia during acute babesiosis. Treatment with T-2 toxin caused a transient significant (P less than 0.05) increase in Babesia parasitemia on day 10 after acute infection and numerical, though statistically nonsignificant, increases in the maximal level and duration of parasitemia during acute babesiosis. Significant (P less than 0.005) recrudescence of parasitemia was observed in the dexamethasone- and cyclophosphamide-treated mice with latent Babesia infection. Treatment with T-2 toxin did not cause recrudescence of parasitemia in mice with latent Babesia infection.     

THE USE OF IMIDOCARB DIPROPRIONATE FOR THE TREATMENT OF THEILERIA SERGENTI INFECTIONS OF CATTLE

SUMMARY Imidocarb diproprionate (IMDP) was used therapeutically at a dose of 1.2 mg IMDP/kg of body weight against Theileria sergenti infections in Hereford heifers on the island of Jeju-Do, Republic of Korea. In the first experiment IMDP was seen to be at least as effective in reducing T. sergenti parasitaemias as primaquine phosphate (primaquine), the drug normally used on the island. In the second and third experiments IMDP was shown to reduce T. sergenti parasitaemias by about 80%. No drug is known to be completely effective against T. sergenti infections but the suggestion is made that IMDP may be used in preference to primaquine for treatment of acutely ill animals since it also has the advantage of eliminating Babesia ovata, another element of the piroplasmosis syndrome on the island.     

A light and electron microscopic study of changes in blood and bone marrow in acute hemorrhagic Trypanosoma vivax infection in calves.

Eleven 6-month-old calves were tsetse fly challenged with a stock of Trypanosoma vivax (IL 2337) that causes hemorrhagic infection. The calves were randomly euthanatized every 4 to 6 days; two other calves served as controls. Peripheral blood changes included anemia, thrombocytopenia, and an initial leukopenia. Later in the course of infection, leukocytosis associated with lymphocytosis and neutropenia developed. Moderate reticulocytosis (highest mean count 3.6 +/- 3.7%, maximum count 9.4%) accompanied the first wave of parasitemia, but poor response (highest mean 0.4 +/- 0.0%) occurred during the second wave, despite the persistence of severe anemia. Light microscopic examination of bone marrow samples showed a drop in the myeloid: erythroid ratio with a decrease in granulocytes, particularly metamyelocytes, bands, and segmenters. Increase in lymphocyte counts corresponded with the appearance of lymphoid nodules within the marrow. Megakaryocytic volume increased significantly in infected animals, and some megakaryocytes showed emperipolesis of red cells, neutrophils, and lymphocytes. Transmission electron microscopic examination of the bone marrow revealed that trypanosomes had crossed the sinusoidal endothelium into the hematopoietic compartment as early as the second day of parasitemia. Macrophages proliferated in the bone marrow; and from the second day of parasitemia until the end of the experimental infection, on day 46, the macrophages had phagocytosed normoblasts, eosinophil and neutrophil myelocytes, metamyelocytes, bands, and segmenters, as well as reticulocytes, erythrocytes, and thrombocytes. Therefore, dyserythropoiesis and dysgranulocytopoiesis were responsible, in part, for the observed anemia and granulocytopenia, respectively.     

The role of IFN-gamma in cattle infected with Theirelia sergenti

The spleen plays an important role in cellular immunity leading to parasite elimination, especially in hemo-protozoan parasite infection. In the present study, we quantified the serum IFN-gamma in splenectomized (SP) and non-splenectomized (NSP) cows infected with Theirelia sergenti (TS) to investigate whether the spleen plays a role in the Th1-type cytokine responses in cows following parasite infection. A transient increase in IFN-gamma was observed in the early stages of infection in the NSP cows, and the cows did not develop parasitemia. In contrast, the SP cows showed no IFN-gamma response at the early stage of infection, and the cows developed parasitemia following anemia. The NSP cows produced IgG1 and IgG2 antibodies to TS-specific, whereas the SP cows showed increases only in IgG1. Increased PBMC proliferation by cocanavalin A stimulation was observed concomitant with the IFN-gamma response. Pretreatment with IFN-gamma suppressed the propagation of TS in the SP cattle. These results suggest that the spleen plays an important role in the resistance to TS infection, leading to a Th1-type immune response. IFN-gamma might contribute to the activation of immunocytes and play an important role in the immune response to resist TS proliferation.     

Babesia ovis infections: Detailed clinical and laboratory observations in the pre- and post-treatment periods of 97 field cases

Ovine babesiosis, caused by Babesia ovis, is of major economic importance in Turkey. The changes in the blood profile of infected animals are informative about the course of infection. The aim of the present study was to evaluate the hematological and biochemical changes in the pre- and post-treatment periods of the natural B. ovis infections. The presence of the parasites was confirmed by microscopy and polymerase chain reaction (PCR) analysis. On the basis of the clinical and laboratory findings, the infections were categorized into different groups according to the degree of anemia and the level of parasitemia. All infected sheep were treated with imidocarb dipropionate (IMDP). The blood pictures in the pre- and post-treatment periods were compared.Pancytopenia occurred in animals with severe anemia and very high parasitemia, and bicytopenia in the other groups. The platelet count (PLT), plateletcrit (PCT) and mean platelet volume (MPV) returned to the normal ranges after treatment, except those in the group with severe anemia. In the biochemical profile, B. ovis infection caused an increase in blood urea nitrogen and total bilirubin, and these parameters returned to normal levels after treatment.The indirect fluorescein antibody test (IFAT) results showed that 38.1% of the cases raised specific antibodies during the period of infection, with titers ranging from 1/160 to 1/640. All of 45 animals re-examined after treatment were seropositive, with high titers that rose up to 1/5120.     

The correlations among serum tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and sialic acids with peripheral lymphocytes in bovine tropical theileriosis

The infection with protozoan parasite Theileria annulata induces changes triggering the activation and/or proliferation of the host lymphocytes. In order to find out the possible correlations among peripheral circulatory lymphocytes, cytokine activities and the level of sialic acids, 50 dairy Holstein cattle, naturally infected with T. annulata, were divided into 4 subgroups according to their parasitemia rates (<1%, 1–3%, 3–5% and >5%). Also, ten non-infected cattle were sampled as control group. Blood samples were taken from jugular vein into acid citrate dextrose-containing tubes for measuring hematological parameters and B and T (CD₄ and CD₈) cell populations and without anticoagulant for TNF-α, IFN-γ and sialic acid concentrations. Remarkable decreases observed in red blood cells (RBCs), white blood cells (WBCs) and packed cell volume (PCV) in infected cattle compared to healthy ones (P < 0.05). Also, with increase in parasitemia rate, total lymphocytes and monocytes alleviated in the diseased groups. By contrast, total neutrohpils and the concentrations of TNF-α, IFN-γ and total sialic acids were significantly elevated (P < 0.05) in infected animals. Accordingly, the circulatory populations of CD₄ and CD₈ T cells and B cells showed a substantial decrease, while a significant increase was observed in T (CD₄ and CD₈) cells in cattle infected with <1% parasitemia rates. Decreased circulatory T cell population shows the ineffective responses of T cells to the stimulatory cytokines such as IFN-γ or TNF-α. On the other hand, the elevation of cytokines (particularly IFN-γ) and sialic acids have presumably an inhibitory role on circulatory B cell population in infected cattle. In addition, a high level of sialic acid concentration indicates the probable role of sialic acid to regulate the parasite-host cell adhesion during sporozoites invasion.     

The post-parturient rise: A comparison of the pattern and relative generic composition of faecal strongyle egg counts in ewes and wethers

Infective larvae of Ostertagia spp. and Cooperia spp. derived from naturally infected dairy calves were subjected to periods of storage of up to 16 weeks at 4°C or 15°C to determine if this treatment would influence their propensity for arrested development in previously worm-free calves. Results showed no significant increase in the propensity of Ostertagia spp. for arrested development in response to the treatments, but a small increase in the case of Cooperia spp.     

奶牛乳房炎主要病原菌感染流行规律研究进展

乳房炎作为全球性奶牛养殖业的重要疾病,因对社会经济、养殖行业和人类健康的巨大影响使人们从发病机理、疾病诊断、预防等方面对其进行了不断的研究与探讨。作者立足于前期尤其是近几年的研究文献,以引起奶牛乳房炎的重要原因——病原菌为中心展开,探讨其感染类型、奶牛与病原菌及病原菌相互之间的关系。奶牛乳房炎是由包括病原菌的侵袭及宿主奶牛自身抵抗力等多种因素共同作用导致,乳腺对病原菌的入侵应答能力,病原与病原之间或促进或抑制都对其发生产生影响。金黄色葡萄球菌是公认的奶牛乳房炎的主要传染性病原菌,其独特的生物学特性使得其在体内难以治疗,并且耐药性普遍存在;无乳链球菌是环境性还是传染性病原尚有争议,在中国感染率较高,对大部分抗生素仍旧敏感,相对于其他病原菌治愈率较高,畜群清除容易;大肠埃希氏菌被作为环境性病原体,乳房炎感染的分离率仍较高,抗生素治疗结果还有待商榷,耐药性普遍存在并增强;牛支原体近十年的感染率有所上升,在国内外都是严重的乳房炎致病因素,治疗效果不佳,且耐药性正在增强,疫苗开发困难。文章对上述几种主要病原菌的流行、危害与防治等研究进行综述,以期为奶牛生产管理和科学研究方面提供参考。     

Alterations of erythrocyte antioxidant mechanisms: antioxidant enzymes, lipid peroxidation and serum trace elements associated with anemia in bovine tropical theileriosis

In order to investigate the alterations of erythrocyte protective antioxidant mechanisms, lipid peroxidation and trace elements associated with anemia in bovine tropical theileriosis, an infected group comprised of 50 crossbred Holstein cattle, about 1-2 years old, naturally infected with Theileria annulata, were divided into 4 subgroups according to their parasitemia rates (<1%, 1-3%, 3-5%, >5%) and also 10 healthy cattle as control were selected. Blood samples were taken and hematological parameters, the activities of antioxidant enzymes including superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase and serum concentrations of some antioxidant trace elements (copper, iron, zinc, manganese and selenium) were measured. As an index of lipid peroxidation, the level of Malondialdehyde (MDA) was also determined. The results showed a conspicuous decrease in the activities of SOD, GPX and catalase (P<0.01), and a significant decrease in the serum concentrations of Cu, Zn, Mn and Se in cattle with higher than 1% parasitemia (P<0.05) compared to the control. In addition, remarkable elevations in the MDA level (P<0.01) and serum concentration of iron (P<0.05) were observed in the infected animals. These findings pointed to the occurrence of exacerbating oxidative injuries to erythrocytes during parasitemia. Furthermore, it can be concluded that infection with T. annulata can interfere with protective antioxidant mechanisms of RBCs against oxidative damages, which promote the development of anemia.     

ORIGINAL ARTICLE: Echinacea purpurea and Allium sativum as immunostimulants in fish culture using Nile tilapia (Oreochromis niloticus)

The study was conducted to evaluate the efficiency of echnicacea (E) and garlic (G) supplemented diets as immunostimulant for tilapia (Oreochromis niloticus). Seven treatments were designed including a control (C). Fish were fed on 35% protein diet at a rate of 3% body weight per day. Echinacea (1.0 ppt) and garlic (3%) were incorporated in the feed, which was administered for periods of 1, 2 and 3 months (summer season), followed by basal diet for 4 more months (winter season). Neutrophil adherence and haematocrit values increased in both supplemented groups with prolonging period of application. The neutrophils adherence was significantly increased in all treatments except group administered echinacea for 1 month. The lymphocytic counts were significantly (p < 0.004) elevated that resulted in a significant increase in the total leucocytic count in groups administered echinacea for 1 and 2 months when compared with the control and/or other treatments. The gain in the body weight and specific growth rate was significantly increased in all supplemented groups (p < 0.004) during summer, but remained without any significant increase after winter. The survival rate was significantly high (>85%) in all the supplemented groups. The percentage of protection, after challenge infection using pathogenic Aeromonas hydrophila was the highest in groups supplemented with echinacea and garlic for 3 months after summer and winter seasons. It could be concluded that echinacea and garlic improve the gain in body weight, survival rate and resistance against challenge infection. Both compounds showed extended effects after withdrawal and improved resistance to cold stress during the winter season. However, a full commercial cost benefit analysis is necessary before recommending their application in aquaculture.