Oxygen uptake by Trypanosoma brucei and there strains of T. vivax

The oxygen uptake of Trypanosoma brucei and three strains of T. vivax was studied in the presence of electron-accepting substrates, α-glycerophosphate or succinate using polarographic electrodes. T. brucei incorporated oxygen only in the presence of α-glycerophosphate. Both a naturally infective (wild strain) and a standard ruminant-passaged strain of T. vivax incorporated oxygen in the presence of α-glycerophosphate and in the presence of succinate. A mouse-adapted strain of T. vivax, however, appeared to be similar to T. brucei in that it incorporated oxygen in the presence of α-glycerophosphate, but did not do so in the presence of succinate.     

Release of Trypanosoma congolense from the microcirculation of cattle by Berenil

Intravenous injection of Berenil (3.5 mg/kg) was followed in 5 min by an increase in the jugular blood parasite count of T. congolense-infected cattle. This increase was statistically significant between 5 and 10 min post Berenil injection. The peak parasitemia reached at 8 min post Berenil was 15 times greater than the pre-treatment parasitemia. It is suggested that the rise in jugular parasitemia results from release into the general circulation of T. congolense organisms which are localized in the microcirculation. Also, because a direct trypanocidal effect of Berenil was not observed in the blood held in vitro, it is suggested that Berenil acts by making trypanosomes available to host defenses, such as the macrophage system.     

Effect of isometamidium on infections by Trypanosoma vivax and T. Evansi in experimentally-infected animals

Assays dealing with the therapeutic and prophylactic activity of isometamidium on experimental infections by Trypanosoma vivax and T. evansi were carried out. The drug was found to be highly effective against T. vivax infection in sheep and cattle in which periods of protection ranging from 118 to 195 days were achieved. No complete effects against infection by T. evansi were observed. The drug was well tolerated in sheep and cattle while side-effects were noted in treated mares. It was concluded that isometamidium could be used to prevent damage and economical losses caused by T. vivax in Venezuela.     

Clinico-pathological and pathomorphological observations in trypanosoma vivax infection cattle

During infection with T. vivax (strain Y 58) four stages can be distinguished: (a) A prepatent period of about 1 week. (b) An acute or fulminating disease characterized by an almost continuous, fluctuating parasitaemia, a decrease of the number of thrombocytes, blood serotonin level, packed cell volume (PCV) and Hb, serum albumin and γ-globulin fractions, a low serum total lipid content and an initial rise with subsequent decrease of the blood bradykinin level. (c) A critical stage of about 4–5 weeks post infection (pi) during which the animal may develop a progressive parasitaemia and may die from disseminated intravascular coagulation. At necropsy microthrombi were a consistent and most significant observation. Accompanying features were oedema and haemorrhages in the various tissues, pulmonary oedema and lobular infarction, patches of necrosis in liver, spleen and adrenals and nephrotic changes in the kidneys. Perivascular mononuclear infiltrates, resulting in local granuloma formation (heart liver), were a constant feature. Related biochemical changes were an increase of the blood urea and serum lactate dehydrogenase levels. (d) A post-critical stage characterized by an ameliorating condition, periods with no or few trypanosomes in the peripheral blood alternating with relatively high peaks of parasitaemia. This was associated with increased blood bradykinin levels, an increase of the PCV and Hb, serum albumin and γ-globulin fractions, number of thrombocytes and blood serotonin level. A return to normal values of the serum total lipids was observed at this stage. Despite this clinical recovery of the animals, at necropsy (on days 90, 98, and 99 pi) there was a moderate to massive histiocyte-lymphocyte-plasma cellular infiltration of the heart, which led to cardiac insufficiency in two heifers.The rise of the d-amino laevulinic acid dehydratase levels of erythrocytes during the decline of the PCV and Hb values is an indication for the production of many young erythrocytes and thereby for the haemolytic nature of the anaemia.     

Increased Inflammatory Mediators in Horses Naturally Infected with Trypanosoma vivax. A Preliminary Study

The aim of this study was to measure the levels of inflammatory mediators in serum from horses naturally infected with Trypanosoma vivax. Banked serum samples collected during a previously reported T. vivax natural infection were used to analyze proinflammatory cytokines such as interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), interleukin 1 (IL-1), interleukin 6 (IL-6), and nitrite/nitrate (NO_x) levels. We evaluated 12 serum samples from horses from a farm in southern Brazil, four of which had parasitological and molecular diagnoses for T. vivax and presented with clinical signs of disease. Cytokines were assessed by quantitative sandwich enzyme-linked immunosorbent assay, and NO_x was measured using the modified Griess method. Levels of IFN-γ, TNF-α, IL-1, IL-6, and NO_x were increased in serum of infected animals compared to that in noninfected animals. Therefore, infection with T. vivax caused an increase in proinflammatory cytokines and nitric oxide content.     

Trypanosoma evansi infection in mainland Spain

An outbreak of Trypanosoma evansi infection that occurred in mainland Spain is described. The outbreak occurred on an equine and camel farm to which dromedary camels from an infected area of the Canary Islands had recently been introduced. One of these camels developed clinical signs and T. evansi was discovered in a blood smear examination. The herd was evaluated in order to determine the extent of the disease. The results showed that 76% of the camels, 35% of the donkeys and 2% of the horses were affected. The animals were isolated and treated using Cymelarsan^® (0.5 mg/kg). After treatment, three blood analysis using parasitological methods revealed negative results. This is the first T. evansi outbreak to have occurred in mainland Spain and the second in mainland Europe, both occurring after the introduction of dromedary camels from the Canary Islands.     

Characteristics of Bacteroides nodosus isolated from cattle

Fourteen isolates of Bacteroides nodosus were cultured from cattle on six farms and examined for colony morphology, pilation, agglutinability, proteolytic activity and pathogenicity for sheep. Where sheep were also present on the farms, isolates from these were compared with those from cattle.Colonies of the bovine isolates were moderately fimbriate. Cells from these colonies were pilated, but not to the extent observed on virulent sheep isolates. The proteolytic activity of the isolates was also less than that described for virulent ovine isolates. When inoculated into sheep, B. nodosus from cattle produced only mild interdigital inflammation. There was no separation of horn characteristic of virulent foot-rot. Preliminary studies, based on agglutination tests, suggest that B. nodosus with different surface antigens may occur in cattle in the same herd and in cattle and sheep on the same farm. On one of six farms isolates from cattle and sheep were indistinguishable by the agglutination test.     

Growth hormones therapy in immune response against Trypanosoma cruzi

Growth hormone (GH) is an important hypophyseal hormone that is primarily involved in body growth and metabolism. In mammals, control of Trypanosoma cruzi parasitism during the acute phase of infection is considered to be critically dependent on direct macrophage activation by cytokines. To explore the possibility that GH might be effective in the treatment of Chagas’ disease, we investigated its effects on the course of T. cruzi infection in rats, focusing our analyses on its influences on parasitemia, NO, TNF-α and IFN-γ concentration and on histopathological alterations and parasite burden in heart tissue. T. cruzi-infected male Wistar rats were intraperitoneally treated with 5 ng/10 g body weight/day of GH. Animals treated with GH showed a significant reduction in the number of blood trypomastigotes during the acute phase of infection compared with untreated animals (P < 0.05). For all experimental days (7, 14 and 21 post infection) of the acute phase, infected and GH treated animals reached higher concentrations of TNF-α, IFN-γ and nitric oxide as compared to untreated and infected counterparts (P < 0.05) Histopathological observations of heart tissue revealed that GH administration also resulted in fewer and smaller amastigote burdens, and less inflammatory infiltrate and tissue disorganization, indicating a reduced parasitism of this tissue. These results show that GH can be considered as an immunomodulator substance for controlling parasite replication and combined with the current drug used may represent in the future a new therapeutic tool to reduce the harmful effects of Chagas’ disease.     

The chemotactic response of murine peritoneal exudate cells to Trypanosoma brucei

A modified Boyden technique was used to assess the chemotactic activity of unstimulated mouse peritoneal exudate cells in the presence of Trypanosoma brucei. Live parasites, the soluble fraction obtained from disrupted trypanosomes, and specific mouse hyperimmune T. brucei sera were unable to generate chemotactic activity. Complexes of whole parasites and hyperimmune sera gave similar control values, whereas significant activity was obtained using a mixture of live trypanosomes and untreated mouse plasma. Immune complexes prepared by pre-incubation of soluble trypanosome fractions with either untreated or heat inactivated (56°C for 30 min) hyperimmune sera, or with mouse plasma, also resulted in enhanced chemotactic activity.     

Cryopreservation of Trypanosoma evansi after DEAE-cellulose purification: Evaluation of infective parameters

Cryopreservation is a method of keeping parasites alive in a laboratory. However, this technique may also damage the parasite. Alternatively, parasites may be maintained by in vitro culture. Unfortunately, for Trypanosoma evansi no effective medium that is able to maintain the parasite for more than 4 months has been described. In this study, we examined the effect of purifying trypomastigote through DEAE-cellulose chromatography before and after cryopreservation, by analyzing the pre-patent period, longevity, parasitemia, and count of viable parasites. Our results showed a three-times increase in the concentration of viable trypomastigote in DEAE-purified cryopreserved parasites as compared to non-DEAE-purified cryopreserved parasites. This indicates that DEAE-cellulose chromatography followed by cryopreservation is an effective method for the storage and preservation of T. evansi, with the advantage that the stocked parasites will be ready to use in molecular biology procedures.     

Occurrence of Trypanosoma evansi in Horses in the State of Minas Gerais,Brazil

Ttrypanosomosis caused by Trypanosoma evansi is a condition that causes significant losses to farmers in endemic areas. This study aimed to report one case of trypanosomiasis in the municipality of Itabira, state of Minas Gerais, Brazil. In November 2010, on a farm with 16 horses, three horses had clinical signs of anemia, limb edema, weight loss, lameness, muscle atrophy, and incoordination of the hind limbs. Trypanosomosis was suspected, and the animals with clinical signs were treated with two doses of diminazene aceturate (7 mg/kg, intramuscular) every 7 days; the treated animals recovered from the clinical symptoms. Blood samples were collected from six horses on the farm, including the three treated animals, to perform polymerase chain reaction specific for T evansi. Four samples were polymerase chain reaction negative, including those collected from three treated horses. However, two other asymptomatic horses were positive for the parasite based on the molecular testing. Based on the results, we concluded that the initial clinical suspect of trypanosomosis was correct, and the treatment used was effective. Probably the disease was introduced to Minas Gerais State through a stallion, which acquired the infection in an endemic area in southern Brazil.     

Agglutination of Brucella abortus cells by sera from cattle experimentally infected with Escherichia coli

Infection of normal adult cattle and sporadic serological Brucella abortus reactors, which no longer had diagnostic titers to B. abortus, with a culture of Escherichia coli isolated from a cow (or its environment) resulted in the production of a primary IgM serum antibody response to both B. abortus and E. coli. Cattle injected with heat killed E. coli and guinea pigs or young calves lacking natural agglutinins to B. abortus injected with live E. coli, produced serum antibody only to E. coli. The subsequent reinjections of live E. coli into the former two groups of cattle resulted in all but one animal in each group producing IgM ‘secondary’ responses to B. abortus of decreasing magnitude, while the anti-E. coli responses increased and eventually switched to synthesis of IgG class antibody. The remaining two animals produced a series of ‘secondary’ responses of IgM antibody to B. abortus similar in amplitude and duration to the primary immune response. The anti-E. coli agglutinins of these two animals increased in titer and IgG class antibody to E. coli was evident after several injections of E. coli. These results indicate that infection of cattle by E. coli can cause a problem in the serological fiagnosis of B. abortus infection. Speculations on the cause of this cross-reaction and ways of minimizing misdiagnosis are discussed.     

The relationship between Theileria taurotragi from eland and Theileria sp. (Idobogo) from cattle

Theileria taurotragi and Theileria sp. (Idobogo) isolated from Kenyan eland and Tanzanian cattle, respectively, have many characteristics in common. It was found that tick-derived stabilites of Theileria sp. (Idobogo) were infective to eland, although only very mild infections developed. Eland that had recovered from Theileria sp. (Idobogo) infections were susceptible to challenge with T. taurotragi stabilate, and similar infections developed in control eland. Cattle that had recovered from Theileria sp. (Idobogo) infection were immune to challenge with T. taurotragi, in contrast to cattle that had recovered from T. taurotragi, which were susceptible to Theileria sp. (Idobogo) challenge. Using T. taurotragi piroplasm antigen in the indirect fluorescent antibody test, a high degree of cross-reaction was observed between T. taurotragi and Theileria sp. (Idobogo) antisera. It would appear that T. taurotragi and Theileria sp. (Idobogo) represent strains of the same species which are adapted to different hosts.     

Purification of cattle oviduct specific proteins and their effect on in vitro embryo development

The purpose of this study was to determine the effect of oviduct specific proteins as a media supplement for in vitro embryo development in cattle. The proteins were extracted from oviducts of cows and precipitated by ammonium sulfate (30%, 40%, 50% and 60%) followed by dialysis in 50 mM Tris–HCl (pH 7.0) buffer. The dialyzed proteins were fractionated into acidic, basic and neutral fractions using SP sephadex cation exchange and DEAE sephadex anion exchange column chromatography respectively. Cow oviduct specific proteins (cOSPs) constituting all the extracted proteins were used as media supplement in three different concentrations (10, 50 and 100 μg/ml) for in vitro maturation, fertilization and culture (IVMFC) of cow oocytes. Acidic, basic and neutral (unbound) fractions were also used as media supplement in three different concentrations (10, 30 and 50 μg/ml) for IVMFC. Cumulus oocytes complexes were collected from slaughterhouse ovaries, washed thoroughly and cultured in maturation media for 24 h in 5% CO₂ at 38.5 °C with maximum humidity. In vitro matured oocytes were co-incubated with in vitro capacitated sperm in Fert-BO media at 38.5 °C for 18 h in 5% CO₂. The fertilized oocytes were washed and cultured in embryo development media for cleavage. After 40–42 h cleavage was observed and embryos were put in the replacement media for further development. The cleavage rates (%) for cOSPs were observed as 68.24±2.46, 69.28±2.05, 61.77±0.93 and 42.62±1.31 at concentrations of 0, 10, 50 and 100 μg/ml respectively. Rates of blastocyst stage development were 14.49±3.61, 21.17±2.77, 14.66±1.06 and 11.98±1.84. These results indicate that addition of cOSP at10 μg/ml increased blastocyst formation as compared to other concentrations (0, 50 and 100 μg/ml). Although acidic, basic and neutral fractions seemed to have no major effect on cleavage rate, but both acidic and neutral fraction of oviduct specific proteins improved the cleavage rate at 30 μg/ml concentration and basic fraction improved the blastocyst formation at 10 μg/ml concentration.     

Molecular detection of Theileria and Babesia infections in cattle

This study was carried out to determine the presence and distribution of tick-borne haemoprotozoan parasites (Theileria and Babesia) in apparently healthy cattle in the East Black Sea Region of Turkey. A total of 389 blood samples were collected from the animals of various ages in six provinces in the region. Prevalence of infection was determined by reverse line blot (RLB) assay. The hypervariable V4 region of the 18S ribosomal RNA (rRNA) gene was amplified with a set of primers for members of the genera Theileria and Babesia. Amplified PCR products were hybridized onto a membrane to which generic- and species-specific oligonucleotide probes were covalently linked. RLB hybridization identified infection in 16.19% of the samples. Blood smears were also examined microscopically for Theileria and/or Babesia spp. and 5.14% were positive. All samples shown to be positive by microscopy also tested positive with RLB assay. Two Theileria (T. annulata and T. buffeli/orientalis) and three Babesia (B. bigemina, B. major and Babesia sp.) species or genotypes were identified in the region. Babesia sp. genotype shared 99% similarity with the previously reported sequences of Babesia sp. Kashi 1, Babesia sp. Kashi 2 and Babesia sp. Kayseri 1. The most frequently found species was T. buffeli/orientalis, present in 11.56% of the samples. T. annulata was identified in five samples (1.28%). Babesia infections were less frequently detected: B. bigemina was found in three samples (0.77%), B. major in two samples (0.51%) and Babesia sp. in five samples (1.28%). A single animal infected with T. buffeli/orientalis was also infected with B. bigemina.     

Effect of zinc supplementation in pregnant mice during experimental Trypanosoma cruzi infection

Zinc is an essential micronutrient and has significant effects on human growth, development, and immune function. Zinc supplementation or deficiency may affect the course of infection. Zinc enhances immune response against a wide range of viral, bacterial, and parasitic pathogens. In the present study, we investigated the effects of zinc sulphate (ZnSO₄) supplementation (20 mg/kg/day) during pregnancy in mice, Swiss Webster strain infected by the Y strain of Trypanosoma cruzi. Oral supplementation of zinc sulphate in pregnant and non-pregnant infected animals did not affect the count of blood parasites as well as tissue parasitism in the heart, liver, and spleen. Zinc supplementation did not alter female body weight, the length of fetuses and neonates, placental size/weight and mortality rate. Among zinc supplied animals, no significant plasmatic zinc concentrations were observed. Concerning to tissue zinc concentrations, only the liver displayed enhanced values as compared to other organs. For placental parasitism, zinc supplied group displayed a significant decrease in amastigote burdens (P < 0.05). However due to the reduced number of parasite burdens in placenta of animals supplied with zinc, these data suggest that zinc was partially effective in up-regulating the host’s immune response against parasite, probably attenuating the infection in fetuses.     

Effect of trypanocides on jugular concentration of Trypanosoma congolense in the West African dwarf sheep

The effects of various trypanocides on parasitaemia was investigated in sheep experimentally-infected with Trypanosoma congolense strain 58/98. Intravenous injection of Berenil at the height of the first parasitaemic wave increased jugular parasite concentration by 12 and 16 times at the 9th and 20th minute post-treatment, respectively. With Pentamidine, maximum counts were 5.0–8.6 times zero-time concentration during the same periods. Peak effects of Samorin, Novidium and Ethidium were observed between the 60th and 90th minutes after drug administration and were 9.5, 6.3 and 3.5 times initial values, respectively. Injection of trypanocides resulted in double peaks of parasitaemia in which the second was usually higher than the first, except with Antrypol and Germani which had no significant effect on parasitaemia. The amplitude, but not the onset of the increase in parasitaemia in sheep, was found to be related to the therapeutic efficacy of the trypanocides in the treatment of Trypanosoma congolense infection in rats.Animals treated with the diamidines (Berenil and Pentamidine) exhibited apparent parasitologic cure of infection in sheep two to four days after treatment. However, administration of any of the drugs one week after the first treatment resulted in flushing of cryptic trypanosomes into the jugular vein and counts as high as 7.63 × 10³ μl^?1 were observed within ten minutes with Berenil. It is suggested that besides their therapeutic use, the diamidines may be of value in the parasitologic diagnosis of sub-patent trypanosomiasis due to Trypanosoma congolense.     

Microscopic analysis of calcium ionophore activated egress of Toxoplasma gondii from the host cell

Toxoplasma gondii invades and destroys nucleated cells of warm blooded hosts in a process which involves several steps: recognition, adhesion, penetration, multiplication inside a parasitophorous vacuole (PV) and egress. The last one is the least understood. Parasite egress from LLC-MK2 cells infected with the RH strain of T. gondii was artificially triggered with 4BrA23187 calcium ionophore. The combination of videomicroscopy, field emission scanning electron microscopy (FESEM), and transmission electron microscopy (TEM) showed that egress does not result from host cell rupture due to overloading with tachyzoites. Videomicroscopy showed that upon calcium ionophore administration parasite rosettes disassemble, the contour of the parasitophorous vacuole disappears and each tachyzoite takes a separate route to the extracellular medium. FESEM and TEM showed the fragmentation of the intravacuolar network, the fragmentation of parasitophorous vacuole membrane and individual tachyzoites with extruded conoids migrating through the cytosol, tightly surrounded by remnants of parasitophorous vacuole membrane or free in the cytosol. Both videomicroscopy and FESEM showed that a single parasite can cross the host cell membrane without disrupting it, while a large number of parasites, egressing simultaneously, rupture the membrane and the cell as a whole. These data suggest that invasion and egress share less similarities than previously believed.     

Non-typhoidal Salmonella encephalopathy involving lipopolysaccharide in cattle

This study assessed the involvement of lipopolysaccharide (LPS) in the non-typhoidal Salmonella encephalopathy (NTSE) caused by a unique isolate of Salmonella enterica serovar Saint-paul (SstpNPG). NTSE was prevented by genetic (deletion of murE) or pharmacologic (polymyxin) disruption of LPS on SstpNPG although the disruption of LPS did not deter brain penetration of the strain. This is the first study to demonstrate that LPS is involved in the manifestations of NTSE.     

Exposure of immunized cattle to prolonged natural challenge of Theileria lawrencei derived from African buffalo (Syncerus caffer)

Cattle were immunized with Theileria lawrencei stabilates either by fortuitous recovery or intentionally by chemoprophylaxis with oxytetracycline. The immunized cattle were exposed together with susceptible control cattle in a paddock where a lethal T. lawrencei challenge derived from two African buffaloes had been established.Theileria lawrencei stabilates used for immunization were of two types: one batch was prepared from Rhipicephalus appendiculatus fed on a buffalo in the paddock, and the other was from a number of tick batches fed on the two buffaloes over a period of 3 months.All the susceptible control cattle exposed in the paddock died of acute Theileria lawrencei infections. Cattle immunized with the composite stabilates survived T. lawrencei challenge for a prolonged period without showing clinical disease. The protection to the T. lawrencei challenge persisted for at least 1.5 years after the composite stabilates had been prepared. The stabilate prepared from ticks fed on a buffalo on one occasion failed to give effective protection since half of these immunized cattle died of T. lawrencei infection when exposed. These results suggest that different immunogenic types of T. lawrencei occur in buffalo which may hinder the effectiveness of a vaccine for T. lawrencei.