A financial analysis of treatment strategies for Trypanosoma evansi in the Brazilian Pantanal

The Brazilian Pantanal is a 138 000 km² tropical seasonal wetland located in the center of South America bordering Bolivia and Paraguay. The Pantanal contains approximately 1100 cattle ranches, 3 million cattle, 49 000 horses and a unique diversity of wildlife. Cattle ranching is the most important economic activity in the Pantanal. This study explores the direct financial impacts of the adoption of seven treatment strategies for the control of Trypanosoma evansi in the Brazilian Pantanal. T. evansi adversely affects the health of the horse population in the region. Horses are indispensable to the cattle ranching industry in the Pantanal. Estimated costs include risk of infection, costs of diagnosis, alternative treatments, collecting animals for treatment, and costs of animal losses. The estimated total cost of T. evansi to the Pantanal region's cattle ranchers is about US$2.4 million and 6462 horses/yr. Results indicate that one preventive and two curative treatment strategies are financially justifiable. The best available technology for the treatment of T. evansi from an economic perspective is a curative treatment employed year-round. This treatment represents an annual net benefit of more than US$2 million or US$1845/ranch and spares about 5783 horses. It represents an annual net benefit of over US$200 000 and 600 horses relative to the currently most widely adopted strategy.     

Prevalence of Surra among camels and horses in Jordan

The prevalence of Trypanosoma evansi infection among camels and horses in Jordan was studied using thick blood smears and inoculation techniques with mice and rats. A total of 437 camels and 83 horses from four climatic zones were surveyed. In addition, 40 donkeys, 32 cattle and 35 goats in contact with infected camels and horses were also tested in the same way. Clinical disease was evident in 8.2% of the camels (36 out of 437) and in 9.6% of the horses (8 out of 83). Infection was limited only to the Sweama area on the Dead Sea (within the warm desert-climatic zone), with prevalence of 30.5% and 33.3%, respectively, for camels and horses. Donkeys, cattle and goats examined were all free from T. evansi. Clinically affected camels were positive by both, thick blood smear and mouse and rat inoculations. Rat and mouse inoculations revealed (X²=3.2, df=1, exact p=0.07) greater number of positive cases in horses than those revealed by thick blood smears. T. evansi-infected camels and horses showed all the clinical signs known for Surra. In addition, it was observed that 100% of infected camels stared at the sun.     

Prevalence of bovine haemoparasites in Aceh province of Northern Sumatra: Implications for imported cattle

A limited seroepidemiological investigation was conducted in Aceh Province to determine the seroprevalence of Anaplasma marginale and Trypanosoma evansi in indigenous cattle and in groups of Sahiwal cross cattle imported from New Zealand in 1985. The results obtained suggest that these parasites are endemic in the areas surveyed; in local cattle, overall prevalence rates of 82% for A. marginale and 61% for T. evansi were obtained.

Thin blood smears were prepared from 42 anaemic adult cattle and 19 clinically normal calves. Babesia bigemina was detected in 5 blood smears and Theileria orientalis in 51. Sahiwal cattle imported from New Zealand suffered high mortalities during their first year in Aceh and it is suggested that imported naive cattle rapidly become infected with blood parasites.     

Chemotherapy of surra in horses and mules with diminazene aceturate

During June–July 2000, an outbreak of surra occurred on an equine breeding farm in Khonkaen Province, Thailand. Forty-two percent of pregnant mares aborted or gave stillbirth and 40% (19/47) of horses and 10% (1/10) of mules died from surra. In August 2000 Trypanosoma evansi were detected in the remaining animals (28 horses and nine mules) on the farm by blood smear and/or the haematocrit centrifuge technique. All animals were treated with diminazene aceturate at 3.5 mg/kg body weight by intramuscular injection on days 0 and 41 of the study. Blood samples of eight randomly selected horses and mules were collected on days 0, 1 and once a week until day 56 and examined for T. evansi by various parasitological techniques. The sera were tested for antibodies against T. evansi using an indirect enzyme linked immunosorbent assay (ELISA).

The results revealed that diminazene aceturate at 3.5 mg/kg appeared to be effective in the first treatment of horses and mules infected with T. evansi. Parasites were cleared from the peripheral blood of horses on days 1 and 7 and mules on days 1 and 14. Thereafter the number of positive animals increased. After the second treatment, 50% of horses and 25% of mules were still positive to surra 24 h after treatment demonstrating that diminazene had no protective effect. Mild to severe toxicity of diminazene was seen in the horses and mules after injection.     

伊氏锥虫伊犁株扩繁及其PFR基因克隆表达和生物信息学分析

【目的】本研究旨在研究锥虫入侵宿主细胞的机制并为其检测方法的建立奠定基础。【方法】采用已分离保存的伊氏锥虫在昆明白小鼠体内进行培养繁殖,对其鞭毛束旁棒(PFR)基因进行克隆,并构建系统发育树。通过生物信息学方法分析和预测PFR蛋白的理化特性、亲疏水性、跨膜区结构、二级结构和三级结构;构建原核表达载体pET28a-PFR,用Western blotting检测PFR重组蛋白的反应原性。【结果】在昆明白小鼠体内成功扩繁伊氏锥虫伊犁株,第5天染虫率达到最高;PFR基因PCR扩增片段大小为834 bp,与冈比亚布氏锥虫PFR基因(XP＿011775815.1)相似性为99.52%,基于PFR蛋白氨基酸序列的进化树显示,该虫株与冈比亚布氏锥虫的亲缘关系也最近。PFR蛋白分子式为C₁₄₁₆H₂₂₈₆N₄₁₆O₄₄₂S₁₁,理论等电点为5.74,是一种碱性、亲水性及不稳定蛋白,没有跨膜区和信号肽,有10个潜在抗原表位;主要定位于细胞质中;PFR蛋白二级结构主要由α-螺旋组成(占92.34%)...     

Drug resistance in Sudanese Trypanosoma evansi

The drug sensitivities of 16 Trypanosoma evansi isolates from Sudan were examined using two different in vitro assays and rodent models. IC₅₀ values (concentration which inhibited incorporation of ³H-hypoxanthine by 50%) obtained in a 40 h assay indicate that most of the isolates were resistant to suramin, a drug which has not been used in Sudan since 1975. Sensitivities for suramin in a 10-day-in vitro assay varied within a 124-fold range. The in vitro results were confirmed by infection/treatment experiments in mice. Sensitivities in vitro for quinapyramine varied within a 166-fold range. In mice, the least sensitive isolates were not cured with dosages up to 10 mg/kg quinapyramine. Based on in vitro results, all isolates appeared to be susceptible to isometamidium.     

An atypical case of Trypanosoma cruzi infection in a young English Mastiff

A case of Trypanosoma cruzi infection in a young English Mastiff from Texas is presented. Clinical signs and laboratory findings included subcutaneous edema, lymphadenopathy, weight loss, and hypoalbuminemia. Cytology of a lymph node revealed numerous amastigotes. No trypomastigotes were observed in buffy coat preparation of peripheral blood, and on histologic evaluation, most organs contained numerous interstitial pseudocysts. Initial serology was positive for both T. cruzi and Leishmania, and immunohistochemistry supported a diagnosis of Leishmania. However, additional serology supported a T. cruzi infection, and cultivation of organisms isolated from a lymph node revealed morphology consistent with T. cruzi. In addition, PCR analysis resulted in a 504 bp fragment with 99% homology to a flagellar protein of T. cruzi. Although uncommon, autochthonous cases of both T. cruzi and Leishmania have been reported in the United States. Clinical signs observed with both diseases can show many similarities, cytology may be indistinguishable, as in this case, and serological cross-reactivity is common. This case demonstrates an unusual presentation of T. cruzi and the use of multiple testing strategies to support its diagnosis.     

A comparative longitudinal study of bovine trypanosomiasis in tsetse-free and tsetse-infested zones of the Amhara Region, northwest Ethiopia

A study was conducted to determine the incidence of trypanosome infections in cattle in tsetse-free and tsetse-infested zones of the Amhara Region of northwest Ethiopia. A total of six sentinel herds were established and the cattle observed during a period of 8 consecutive months. The prevalence of seropositive cattle was high in both the tsetse-free and tsetse-infested zones. The average monthly incidence of trypanosome infection, determined using molecular diagnostic tools, was 20.9% and 25.7% in the tsetse-free and the tsetse-infested zones, respectively. In the tsetse-free, Trypanosoma vivax was responsible for 90.9% of the cattle trypanosome infections. In the tsetse-infested zone, Trypanosoma congolense and T. vivax contributed almost equally to the trypanosome infections in cattle. Trypanosome infection, regardless of species, resulted in anaemia as evidenced by a significant decrease in the packed cell volume of the infected animal. The outcome of this longitudinal study suggests that control of trypanosomiasis in the Amhara Region cannot be achieved by tsetse control alone. Supplemental measures to include drug therapy and biting fly control are discussed.     

Comparative studies on N''Dama and zebu cattle following repeated infections with Trypanosoma congolense

Twenty N'Dama and eight zebu cattle were inoculated intradermally with bloodstream forms of a cloned strain of Trypanosoma congolense originating from East Africa. All inoculated cattle became parasitaemic. Zebus showed consistently higher levels of parasitaemia and lower packed red cell volume (PCV) percentages than did N'Damas. Three of the eight zebus required treatment when high numbers of trypanosomes were present in the blood and PCV values dropped below 15 per cent. None of the N'Dama cattle needed treatment. Statistical analysis was performed on the data to assess the variability of parasitaemia and PCV levels before and during infection of the N'Dama cattle. The variation in PCV values was large between individuals during the early stages of the disease and diminished as infection continued. After trypanocidal drug treatment and a recovery period of 14 months, the same animals were inoculated intradermally with T congolense bloodstream forms isolated and cloned in The Gambia. Differences in susceptibility to the ensuing disease were apparent when comparing N'Dama and zebu cattle. Five zebu cattle needed trypanocidal drug treatment, while none of the N'Damas needed drug intervention. Ranking the 20 infected N'Damas according to average PCV levels revealed that the animals responded similarly to both infections.     

Epidemiology of theilerioses in the Trans-Mara division, Kenya: Husbandry and disease background and preliminary investigations on theilerioses in calves

All the calves born (116) into 3 Maasai cattle herds in the Trans-Mara Division of Kenya, between August 1978 and October 1979, were recruited into a monthly health study which concentrated on theileriosis. Twenty-two of the calves died before they were 6 months of age, but the mortality only increased to 25% by the time the calves reached 18 months of age. The mean birth weight of calves was 17.5 kg while at 190 days post-birth the mean weight was 53.4 kg. The main causes of mortality were starvation (7.8%), neonatal diarrhoea (2.6%), chronic indigestion (2.6%) and theileriosis (2.6%) due to Theileria parva and T. mutans infections. The calves were infected with ticks from birth (mostly Rhipicephalus appendiculatus and Amblyomma spp.) and the first Theileria schizonts were detected on Day 17 post birth and reached a maximum of 18.4% of calves in the 11th week post-birth. Seasonal peaks of macroschizont incidence occurred in February and July. All calves had patent Theileria piroplasm infections by the time they were 5 months old and 44% had shown patent Theileria macroschizont infections by 6–7 months of age. Generally low parasitosis of Theileria piroplasms and schizonts occurred. Serology using the indirect fluorescent antibody test showed a high proportion of calves received antibodies against T. mutans and T. parva from their dams by way of colostrum. The majority of calves also had active antibody responses against T. mutans and T. parva by the time they were 6 months of age. There was a correlation between the pre-patent period of piroplasms and active antibody responses to T. mutans and between the prepatent period of schizonts and an antibody response to T. parva. Eighteen older calves developed T. velifera infections. “Turning sickness” due to Theileria infection in the brain was detected in older cattle. Other blood parasites such as Trypanosoma vivax, T. congolense, Anaplasma marginale and Babesia bigemina occurred at patent levels at a lower incidence than Theileria spp. and did not cause disease problems in the calves. The calf population was highly resistant to theileriosis since they had a 100% morbidity, but only 2.6% mortality. Theileria infections would appear to have an important effect on the growth of calves but this and many aspects of the epidemiology of theileriosis in the area required more intensive sampling.     

Zinc supplementation increases resistance to experimental infection by Trypanosoma cruzi

It is well recognized that zinc is an essential trace element for all organisms, influencing growth and affecting the development and integrity of the immune system. It is also well known that the protective response against Trypanosoma cruzi depends on both innate and acquired immunity and for the control of the parasite load and host survival, the participation of special cells such natural killer (NK), T and B lymphocytes and macrophages are required. So the aims of this study were to evaluate the effects of zinc supplementation on the host's immune response infected with T. cruzi. Our data point in the direction that zinc supplementation triggered enhanced thymocyte and splenocyte proliferation as compared to unsupplied group of animals. It is also important to emphasize that interleukin-12 (IL-12) participates in the resistance to several intracellular pathogens including T. cruzi. Our findings demonstrate an enhanced production of IL-12 during the acute phase of infection in zinc-supplied groups. So we conclude that zinc supplementation leads to an effective host's immune response by up-modulating the host's immune response, thus contributing in the reduction of blood parasites and the harmful pathogenic effects of the experimental Chagas’ disease.     

Differences in cloning and sub-cloning success rates in four stocks of Trypanosoma evansi and variation in suramin resistance of the clones

Four Trypanosoma evansi stocks with sensitivity to suramin in mice ranging from 0.05 to 160 mg kg⁻¹ were cloned and sub-cloned and the sensitivity of the clones determined. The results suggest that it is easier to clone and sub-clone trypanosome stocks which are sensitive to suramin than those that are resistant to the action of the drug. The clones obtained from the four stocks had sensitivities to suramin which were similar to or different from the parent stocks. These results are important in view of the development of resistance for, in the presence of suramin, these resistant yet heterogeneous populations would provide the material from which selective processes could operate. These observations also suggest that the maintenance and spread of suramin-resistant trypanosomes might be curtailed by their comparative inability to establish themselves in a new host.     

犬弓首蛔虫感染比格犬不同阶段肝circRNAs表达模式的分析

旨在探究终末宿主肝环状RNA （circRNAs）在犬弓首蛔虫诱导致病过程中的潜在调控作用。18只6~7周龄的比格犬被平均分为3组（感染后0.5 d组、感染后1 d组和感染后36 d组），每组包括感染和对照各3只，分别于感染后对应的时间点收集肝样品。提取感染和对照组肝的总RNA，利用高通量RNA测序，构建circRNA文库，对差异表达的circRNAs进行分析，并对差异表达circRNAs的亲本基因进行GO注释和KEGG富集分析。随机选取9个差异表达circRNAs进行实时荧光定量PCR验证。结果显示，与对照组相比，在感染后0.5、1和36 d分别有94、103和84个犬肝的差异表达circRNAs。Venn图显示，3个感染阶段没有共同表达的差异circRNAs；GO和KEGG分析显示，差异表达的circRNAs参与了宿主肝免疫与炎症的相关通路，其中差异表达的circRNA novel_circ_0016108、novel_circ_0016184和novel_circ_0027468与犬弓首蛔虫引起的天然免疫相关；novel_circ_0002212参与了犬弓首蛔虫引起的肝炎症反应；novel_circ_0002212和novel_circ_0019907在宿主肝抵抗犬弓首蛔虫侵袭的过程中发挥作用。RT-qPCR验证结果显示大多数差异表达的circRNAs与高通量测序结果一致。结果表明，肝circRNAs在犬弓首蛔虫感染终末宿主的过程中发挥着重要作用，这为进一步探究犬弓首蛔虫与宿主之间的相互作用提供了新的信息，并且也为促进疾病干预措施的制定提供参考。     

Antigenic alteration in major piroplasm surface proteins of Theileria sergenti during infection

Theileria sergenti piroplasms were purified from different parasitemia peaks of cattle infected with parasitized erythrocytes or sporozoites during persistent infection. Their activities with monoclonal antibodies 13F5 and C9, which recognize 23 kDa and 32 kDa piroplasm surface proteins, respectively, were analyzed. Antigenic differences were observed among parasites from different parasitemia peaks during persistent infection when cattle were infected with sporozoites. Results of two-dimensional polyacrylamide gel electrophoresis showed that the 23 and 32 kDa proteins were expressed in all samples tested, regardless of their reactivities with the monoclonal antobodies. In contrast, parasites obtained from cattle inoculated with parasitized erythrocytes showed no antigenic alteration over a 2 month observation period. The results suggest that antigenic alteration of T. sergenti during persistent infection is related to whether the parasites proliferate through extraerythorocytic schizont stage in cattle or sporozoite and other sexual stages in tick vector.     

Antibody responses to Toxoplasma gondii antigens in aqueous and cerebrospinal fluids of cats infected with T. gondii and FIV.

Antibodies to antigens of Toxoplasma gondii were measured in the aqueous and cerebrospinal fluid (CSF) of 16 specific-pathogen free kittens experimentally infected with feline immunodeficiency virus (FIV), T. gondii, or both pathogens. The results indicated that all cats infected with T. gondii had antibody responses to antigens of T. gondii in both aqueous fluids and CSF. Co-infection with FIV did not affect antibody levels. Aqueous fluids from eyes of cats with toxoplasmic retinochoroiditis did not necessarily have higher antibody levels than those from eyes without lesions. Antibodies to T. gondii were also detected in the CSF of two cats from whose brains no parasites were isolated by in vivo mouse inoculation. Total IgG did not increase significantly in the aqueous fluids and CSF of cats infected with T. gondii whether or not they were also infected with FIV.     

The effect of T. vivax infection in west African dwarf goats on energy and nitrogen metabolism.

To investigate how T. vivax affects metabolism in dwarf goats, nine wethers (infection group) given alfalfa pellets ad libitum were infected intravenously and food intake was recorded up to 49 days after infection in the infection group and in the control group (n = 9). Controls received the same diet, ad libitum before infection and in restricted amounts after infection in order to obtain similar intakes in the two groups. Digestible organic matter intake (DOMI) and nitrogen balance (NB) were determined during four balance trials. All animals were bled regularly to measure parasitaemia, packed cell volume (PCV) and a number of serum metabolites. All infected animals showed symptoms typical for T. vivax infection as judged by parasitaemia, PCV and rectal temperature. Infection had a non-uniform negative effect on food intake. Compared with controls at equal DOMI, NB was lower in infected animals, the difference being significant 4 weeks after infection. This was caused by a gradual increase in NB at equal DOMI of the control group. The NB of the ad libitum fed infected animals 2 and 4 weeks after infection was comparable to values normally found in healthy ad libitum fed dwarf goats with an equal DOMI. NEFA values in serum were significantly elevated after infection. Except for two infected animals with an extremely low food intake towards the end of the experiment, no rise in serum ketone bodies was evident. After infection, serum protein increased, differences with controls being significant 4 and 7 weeks after infection.(ABSTRACT TRUNCATED AT 250 WORDS)     

Equine trypanosomosis in the Central River Division of The Gambia: a study of veterinary gate-clinic consultation records

The objective of this study was to provide epidemiological information of equine trypanosomosis in the Central River Division (CRD) of The Gambia. Therefore, 2285 consultations records of equines, admitted in a gate-clinic at Sololo in CRD, were studied retrospectively. The data were recorded in the period between September 1995 and July 2002 and comprised consultations of 2113 horses and 172 donkeys.

‘Trypanosome infection’ was the most frequently diagnosed condition and accounted for 61% of the cases. Horses were more frequently diagnosed with trypanosome infections than donkeys (p < 0.001), with an occurrence of 63% compared to 43% in donkeys. In both horses and donkeys, trypanosome infections were mainly due to Trypanosoma congolense (64%) and T. vivax (32%). There was no difference observed in the occurrence of trypanosome infections in male or female donkeys (p = 0.585), but there were more female (67.8%) horses observed with trypanosome infections than male horses (60.7%; p = 0.003). There was no difference observed in the occurrence of trypanosome infections in donkeys older or younger than 1 year (p = 0.130), but more older horses (63.2% >1 year) were observed with trypanosome infections than young horses (54.5% <1 year; p = 0.033).

The number of donkeys and horses with trypanosome infections decreased during the rainy season (June–September).

The majority of equines that were admitted with trypanosome infections were severely anaemic. The average packed cell volume (PCV) declined with increasing parasitaemia (p = 0.006).

Seventy-four percent of the farmers’ predictions of trypanosome infections in their equines were confirmed by darkground-microscopy. That proved that farmers had a fairly accurate knowledge of the diseases affecting their equines.

The treatments executed at the gate-clinic were generally effective. The few (0.4%) relapses of the T. vivax infections that were previously treated with diminazene aceturate in this study were not sufficient to prove drug resistance.

The study showed that the analysis of consultation records at a gate-clinic can provide complementary information to conventional epidemiological studies in the same research area.     

中国不同地区羊吕氏泰勒虫分子流行病学调查及遗传进化分析

为了解吕氏泰勒虫在中国不同地区山羊和绵羊中的流行情况,本研究应用基于吕氏泰勒虫18S rRNA基因位点的PCR检测方法,对采自中国河南、甘肃、陕西、山西、贵州、云南、新疆7个地区的281份羊血液样品进行检测,并对阳性样品测序以进行序列分析。结果显示,羊吕氏泰勒虫总感染率为35.23%（99/281）。不同采样点羊吕氏泰勒虫感染率差异极显著（P<0.01）,其中河南省羊吕氏泰勒虫感染率最高（98%,49/50）,新疆最低（0）。绵羊和山羊吕氏泰勒虫感染率分别为51.67%（31/60）和30.77%（68/221）,差异极显著（P<0.01）;放牧羊吕氏泰勒虫感染率（41.55%,86/207）极显著高于舍饲羊（17.57%,13/74）（P<0.01）;羊吕氏泰勒虫感染率在春、夏、秋及冬四季分别为56.00%（28/50）、42.75%（59/138）、9.43%（5/53）和17.50%（7/40）,差异极显著（P<0.01）;≥ 12月龄和<12月龄羊吕氏泰勒虫感染率分别为33.50%（67/200）和39.51%（32/81）,差异不显著（P>0.05）。此外,遗传进化分析表明,本研究获得的羊吕氏泰勒虫分离株与中国流行的吕氏泰勒虫分离株同源性高达99.60%以上,且位于同一分支上。本研究为进一步了解中国不同地区羊吕氏泰勒虫的流行现状及分布提供了重要参考依据。     

Adaptation and validation of antibody-ELISA using dried blood spots on filter paper for epidemiological surveys of tsetse-transmitted trypanosomosis in cattle

The indirect enzyme-linked immunosorbent assay (ELISA) for the detection of anti-trypanosomal antibodies in bovine serum was adapted for use with dried blood spots on filter paper.

Absorbance (450 nm) results for samples were expressed as percent positivity, i.e. percentage of the median absorbance result of four replicates of the strong positive control serum.

The antibody-ELISA was evaluated in Zambia for use in epidemiological surveys of the prevalence of tsetse-transmitted bovine trypanosomosis. Known negative samples (sera, n=209; blood spots, n=466) were obtained from cattle from closed herds in tsetse-free areas close to Lusaka. Known positive samples (sera, n=367; blood spots, n=278) were obtained from cattle in Zambia's Central, Lusaka and Eastern Provinces, diagnosed as being infected with Trypanosoma brucei, T. congolense, or T. vivax using the phase-contrast buffy-coat technique or Giemsa-stained thick and thin blood smears. For sera (at a cut-off value of 23.0% positivity) sensitivity and specificity were 86.1 and 95.2%, respectively. For bloodspots (at a cut-off value of 18.8% positivity) sensitivity and specificity were 96.8 and 95.7%, respectively. The implications of persistence of antibodies following treatment or self-cure are discussed.     

Evaluation of loop-mediated isothermal amplification (LAMP), PCR and parasitological tests for detection of Trypanosoma evansi in experimentally infected pigs

Six surra negative piglets (6-week-old) were infected with Trypanosoma evansi and two uninfected piglets were used as negative controls. Detection performances of various diagnostic tests (LAMP, PCR and parasitological tests) were compared by analysing blood samples collected weekly over a period of 11 weeks. With a two by two analysis without a gold standard, all methods were 100% specific. MI had the highest sensitivity of 65%, while LAMP, PCR, MHCT and TBS had sensitivities of 45, 33, 38 and 24%, respectively. However, when the analysis was done using MI as a gold standard, the sensitivity of MHCT was the highest at 53% followed by LAMP, PCR and TBS at 49, 44 and 35%, respectively. All methods gave high specificity above 60%. This study validates LAMP as an alternative method for the diagnosis of surra.