Detection of antibodies in Trypanosome-infected cattle by means of a microplate enzyme-linked immunosorbent assay

Summary A micromodification of the enzyme-linked immunosorbent assay (ELISA) was evaluated for its potential application in the immunodiagnosis of bovine trypanosomiasis. Serum samples from infected and non-infected Zebu cattle and samples from Friesian cattle with experimental infections were examined for the presence of trypanosomal antibodies. There were significant differences between the microELISA values obtained with samples from infected and non-infected cattle. During the course of infection microELISA values were found to fluctuate and the antibody response varied in individual animals. The test did not distinguish between infections withTrypanosoma brucei, T. vivax andT. congolense. There were no cross-reactions between trypanosome antigens and serum samples from cattle infected withT. theileri, Theileria parva, Th. mutans, Th. annulata, Babesia divergens andAnaplasma marginale.     

Parasite kinetics and immune responses in efferent prefemoral lymph draining skin reactions induced by tsetse-transmitted Trypanosoma congolense

Localised skin reactions (chancres) occurred on the flanks of cattle at the sites of deposition by tsetse flies of metacyclic forms of Trypanosoma congolense. Marked enlargement of the draining prefemoral lymph nodes accompanied the development of the skin reactions. Lymph from these nodes was collected through polyethylene cannulae inserted into the efferent lymphatics, and examined for trypanosomes, cells and antibody content. Within 6-9 days after infected tsetse fly bite, trypanosomes were detected in the efferent lymph; this preceded their appearance in the blood by 3-6 days, indicating that the lymphatic system acted as a major route for the passage of trypanosomes from the skin into the bloodstream. Responses induced in the draining lymph node as a result of trypanosome migration included a 2-3-fold increase in the volume of lymph and up to a 10-fold increase in lymphocyte output, including blast lymphocytes and plasma cells. Neutralising antibodies to metacyclic trypanosomes were detected in lymph and serum by Day 14 after infection, although in 2 out of 4 animals investigated, they were not demonstrated in serum until Day 18. Trypanosomes were also found in small numbers in efferent lymph of the prefemoral lymph node on the flank contralateral to the infected tsetse bites after development of parasitaemia. Increases in lymph flow and cellular output occurred about the same time in the ipsilateral and the contralateral efferent lymphatics, but were significantly less in the latter. Homologous challenge of immunised calves with tsetse-transmitted parasites revealed that trypanosomes were eliminated at the level of the skin or within the draining lymph node, as no parasites were detected in efferent lymph.     

Apparent lack of a domestic animal reservoir in Gambiense sleeping sickness in northwest Uganda

The role played by domestic animals in the transmission of gambiense Human African Trypanosomosis remains uncertain. Northwest Uganda is endemic for Trypanosoma brucei gambiense. Of the 3267 blood samples from domestic animals in four counties examined by hematocrit centrifugation technique (HCT), 210 (6.4%) were positive for trypanosomes. The prevalence of animal trypanosomosis was estimated at 13.8% in Terego County, 4.2% in East Moyo County, 3.1% in Koboko County, and zero in West Moyo County. The trypanosome infection rates varied from 0.2% in goats, 3.5% in dogs, 5.0% in sheep, 7.5% in cattle, to 15.5% in pigs. DNA was extracted from the blood samples by Chelex method, Sigma and Qiagen DNA extraction Kits. A total of 417(12.8%) DNA samples tested positive by polymerase chain reaction (PCR) using T. brucei species specific primers (TBR) indicating that the DNA was of Trypanozoon trypanosomes while 2850 (87.2%) samples were TBR-PCR negative. The T. brucei infection rates based on TBR-PCR were highest in pigs with 21.7%, followed by cattle (14.5%), dogs (12.4%), sheep (10.8%), and lowest in goats with 3.2%, which indicated that pigs were most bitten by infected tsetse than other domestic animals. TBR-PCR detected 6.3% more infected domestic animals that had been missed, and confirmed the 6.4% cases detected by HCT in the field. Statistical analysis done using one-way ANOVA Kruskal-Wallis test (Prism version 5.0) showed no significant difference in trypanosome infections among domestic animals using both HCT and TBR-PCR techniques in the different counties (Confidence Interval of 95%, p-values >0.05). All the 417 trypanosome DNA samples were negative by PCR using two sets of primers specific for the T. b. gambiense specific glycoprotein gene and serum resistance associated gene of T. b. rhodesiense, indicating that they were probably not from the two human infective trypanosomes. Polymerase chain reaction using primers based on ribosomal internal transcribed spacer-1 region (ITS-PCR) resolved the 417 DNA of trypanosome samples into 323 (77.5%) as single trypanosome infections due to T. brucei and 39 (9.4%) mixed infections but missed detecting 55 (13.1%) samples, possibly because of the low sensitivity of ITS-PCR as compared to TBR-PCR. The 31 mixed infections were due to T. brucei (T.b) and T. vivax (T.v); while 8 mixed infections were of T. congolense (T.c) and T. brucei but no mixed trypanosome infections with T. congolense, T. brucei, and T. vivax were detected. Statistical analysis done using one way ANOVA Kruskal-Wallis test (Prism version 5.0) to compare single and mixed trypanosome infections showed no significant difference in trypanosome infections due to single (T.v, T.b, T.c) and mixed (T.v+T.b; T.v+T.c; T.b+T.c; T.v+T.b+T.c) trypanosome species among domestic animals in the different counties using ITS-PCR technique (Confidence Interval of 95%, p-values >0.05). It was concluded that domestic animals in northwest Uganda were probably not reservoirs of T. b. gambiense and there was no infection, as yet, with T. b. rhodesiense parasites.     

Relationships between trypanosome infection measured by antigen detection enzyme immunoassays, anaemia and growth in trypanotolerant N'Dama cattle.

Relationships were evaluated between trypanosome infection as measured by antigen detection enzyme immunoassays (antigen ELISA), anaemia as determined by average packed red cell volume (PCV), and animal performance as assessed by daily weight gain in 99 N'Dama cattle in Gabon exposed to natural tsetse challenge at 11.5 months of age and recorded 14 times over a 13 week period. Approximately half the animals were found to be infected for an average of five of the 14 times that they were examined: 38% with Trypanosoma congolense, 13% with Trypanosoma vivax and 49% with a mixed infection. Trypanosoma congolense infections had significant deleterious effects on animal growth, while T. vivax infections did not. Animals found on several occasions to be infected with T. congolense had significantly lower PCV values than those demonstrated to be infected on fewer occasions. No relationship was found between mean optical density (OD) values in antigen ELISA and PCV values. Animals capable of maintaining PCV values, even when antigen ELISA positive on a high number of occasions, grew at the same rate as uninfected animals. Animals that could not maintain PCV values when infected had poorer growth. Antigen ELISA has the potential to increase the efficiency of selection of trypanotolerant N'Dama cattle under tsetse challenge in the field, in three main ways. (1) Accurate identification of trypanosome species, especially in mixed species infections, clarifies relations between infection, anaemia and animal performance. (2) Detection of animals antigenaemic without patent parasitaemia could allow individuals with superior ability to control trypanosome infection to be identified. (3) More accurate measurement of the proportion of time an animal is infected allows more accurate evaluation of its anaemia control capability.     

Larval migration inhibition activity in abomasal mucus and serum from calves infected with Ostertagia ostertagi

The present study investigated whether abomasal mucus from calves naturally infected with gastrointestinal nematodes possessed larval migration inhibition (LMI) activity in vitro, and whether LMI activity was greater in mucus from previously immunised animals, compared to primary infected and uninfected calves. LMI activity was also assessed in serum from calves during both natural and artificial Ostertagia ostertagi infections, in an attempt to monitor the development of acquired immunity. Both abomasal mucus and serum exhibited larval paralysing activity. Although the LMI capacity of the abomasal mucus was very variable, the highest paralysing activity was consistently observed in mucus from previously immunised calves. LMI activity in serum increased significantly during both artificial and natural Ostertagia infections. After a challenge infection, sera from immunised animals showed a significantly higher LMI capacity, compared to previously uninfected calves. Moreover, serum LMI activity was significantly negatively correlated with Ostertagia worm counts after the challenge infection. The present results suggest that LMI activity in serum and/or abomasal mucus reflects a protective immune response against O. ostertagi in the abomasal mucosa.     

Isometamidium chloride prophylaxis against Trypanosoma congolense challenge and the development of immune responses in Boran cattle

Twenty-four Boran cattle were injected with isometamidium chloride (1 mg/kg bodyweight) to investigate the duration of drug-induced prophylaxis against infection by metacyclic forms of Trypanosoma congolense and to determine if specific antibody responses to the organism were mounted by animals under chemoprophylactic cover. Complete protection against either single challenge by five tsetse flies infected with T congolense, or repeated challenge at monthly intervals by five tsetse flies, lasted for five months. Six months after treatment, two-thirds of the cattle were resistant to challenge, irrespective of whether subjected to single or multiple challenge with trypanosome-infected tsetse flies, or titrated doses of in vitro-cultured metacyclic forms of T congolense (5 X 10(2) to 5 X 10(5) organisms), inoculated intradermally. No animal which resisted infection developed detectable skin reactions at the site of deposition of metacyclic trypanosomes or produced trypanosome-specific antibodies. It was concluded that drug residues effectively limited trypanosome multiplication at the site of deposition in the skin, thus preventing subsequent parasitaemia or priming of the host's immune response.     

Non-immune control of trypanosomosis: in vitro oxidative burst of PMA- and trypanosome-stimulated neutrophils of Boran and N'Dama cattle

An in vitro assay that measures the generation of superoxide anions (O2-) was used to assess the level of oxidative burst of phorbol myristate acetate (PMA)- and trypanosome-stimulated neutrophils isolated from healthy Boran and N'Dama cattle, and those infected with Trypanosoma congolense. PMA stimulation of healthy bovine neutrophils resulted in between 300-400 % increase in O2- generation. Neutrophils of Boran cattle exhibited slightly higher but insignificant O2- generation capacity than those of the N'Dama breed. In vitro stimulation by trypanosomes of neutrophils isolated from Trypanosoma congolense-infected cattle caused significant increases in O2- generation, especially on days 14, 28 and 42 post-infection, of both breeds of cattle. No significant differences were observed in O2- generation capacity of the neutrophils of both breeds of infected cattle throughout the period of assay. The results of this study have shown that PMA and trypanosomes do cause an enhanced in vitro oxidative burst, hence trypanosome phagocytosis and killing activity of neutrophils. Neutrophils have been shown to play very significant roles in parasite clearance, hence reduction of trypanosome parasitaemia. The rates of both in vitro generation of O2- and trypanosome phagocytosis over time did not differ significantly between Boran and N'Dama breeds of cattle, even during T congolense infection in this study. Hence, it may be inferred that sustained and higher parasitaemia, more pronounced neutropenia, inadequate bone marrow response and less effective trypanosome-specific immune response, rather than defective neutrophil trypanosome destruction, may be the problem of trypanosusceptible cattle breeds.     

The application of PCR-ELISA to the detection of Trypanosoma brucei and T. vivax infections in livestock

Teneral tsetse flies infected with either Trypanosoma brucei or T. vivax were fed on healthy cattle. Blood samples collected daily from the cattle were examined by microscopy for the presence of trypanosomes, in thick smear, thin smear and in the buffy coat (BC). All the cattle fed upon by infected tsetse developed a fluctuating parasitaemia. DNA was extracted from the blood of these cattle and subjected to polymerase chain reaction (PCR) using oligonucleotide primers specific for T. brucei or T. vivax. The PCR products unique to either T. brucei or T. vivax were identified following amplification of DNA from the blood samples of infected cattle, whereas none was detectable in the DNA from the blood of the cattle exposed to non-infected teneral tsetse. In a concurrent set of experiments, one of the oligonucleotide primers in each pair was biotinylated for use in PCR-ELISA to examine all the blood samples with this assay. Both the PCR and the PCR-ELISA revealed trypanosome DNA in 85% of blood samples serially collected from the cattle experimentally infected with T. brucei. In contrast, the parasitological assays showed trypanosomes in only 21% of the samples. In the blood samples from cattle experimentally infected with T. vivax, PCR and PCR-ELISA revealed trypanosome DNA in 93 and 94%, respectively. Microscopy revealed parasites in only 63% of the BCs prepared from these cattle. Neither PCR nor PCR-ELISA detected any trypanosome DNA in blood samples collected from the animals in the trypanosome-free areas. However, both assays revealed the presence of trypanosome DNA in a number of blood samples from cattle in trypanosomosis-endemic areas.     

Immunosuppressive effect of Trypanosoma congolense and Trypanosoma vivax on the secondary immune response of cattle to Mycoplasma mycoides subsp mycoides.

Trypanosoma congolense and T vivax infections in cattle were shown to have a suppressive effect on the secondary humoral immune response to M mycoides. The trypanosome infections caused an involution of the thymus. The secondary response was biphasic in nature and was unrelated to infection with trypanosomes.     

Analysis of host genetic factors influencing African trypanosome species infection in a cohort of Tanzanian Bos indicus cattle

Trypanosomosis caused by infection with protozoan parasites of the genus Trypanosoma is a major health constraint to cattle production in many African countries. One hundred and seventy one Bos indicus cattle from traditional pastoral Maasai (87) and more intensively managed Boran (84) animals in Tanzania were screened by PCR for the presence of African animal trypanosomes (Trypanosoma congolense, Trypanosoma vivax and Trypanosoma brucei), using blood samples archived on FTA cards. All cattle screened for trypanosomes were also genotyped at the highly polymorphic major histocompatibility complex (MHC) class II DRB3 locus to investigate possible associations between host MHC and trypanosome infection. Overall, 23.4% of the 171 cattle tested positive for at least one of the three trypanosome species. The prevalence of individual trypanosome species was 8.8% (T. congolense), 4.7% (T. vivax) and 15.8% (T. brucei). The high prevalence of T. brucei compared with T. congolense and T. vivax was unexpected as this species has previously been considered to be of lesser importance in terms of African bovine trypanosomosis. Significantly higher numbers of Maasai cattle were infected with T. brucei (23.0%, p=0.009) and T. congolense (13.8%, p=0.019) compared with Boran cattle (8.3% and 3.6%, respectively). Analysis of BoLA-DRB3 diversity in this cohort identified extensive allelic diversity. Thirty-three BoLA-DRB3 PCR-RFLP defined alleles were identified. One allele (DRB3*15) was significantly associated with an increased risk (odds ratio, OR=2.71, p=0.034) of T. brucei infection and three alleles (DRB3*35, *16 and *23) were associated with increased risk of T. congolense infection. While further work is required to dissect the role of these alleles in susceptibility to T. brucei and T. congolense infections, this study demonstrates the utility of FTA archived blood samples in combined molecular analyses of both host and pathogen.     

Observations on Trypanosoma theileri Infection in Cattle

Naturally occurring Trypanosoma theileri infection was studied in two cattle herds. Herd A was a dairy herd of approximately 250. Herd B was an isolated herd of 32 and contained both dairy and beef breeds. Blood samples were collected from all animals in Herd A during July and August on two successive years. Samples were collected from Herd B at monthly intervals. Total leukocyte and differential counts packed cell volume determinations, and trypanosome cultures were made on each sample.

Infection was detected in all age groups between seven months and fifteen years but it was rare in calves. Infected animals were not consistently positive for trypanosomes on consecutive blood cultures and there was considerable variation between infected individuals. Positive cultures were usually obtained from some animals while others were positive intermittently. No correlation was found between trypanosome isolations and the season of the year.

A correlation was found between trypanosome isolation and lymphocytosis. Of the 920 blood samples examined, approximately one in every five trypanosome positive samples had lymphocyte levels in the Bendixen positive range. Approximately one in every twenty trypanosome negative samples had lymphocyte numbers in the Bendixen positive range. Evidence indicated that trypanosome isolation from animals with lymphocytosis was not caused by increased numbers of infected buffy coat cells in the inoculum cultured.

Eight calves were inoculated intravenously with trypanosome-infected blood. Lymphocyte numbers increased an average of 3549 per cumm above pre-inoculation levels in seven and remained essentially unchanged in one. Prior to inoculation with infective blood, two of the calves were intravenously inoculated with trypanosome-infected blood that had been frozen and thawed to kill the trypanosomes contained in it. Neither developed lymphocytosis following this inoculation.

No clinical disease problems which could be attributed to trypanosome infection were found.

     

Trypanosome infection rate in cattle at Nguruman, Kenya

Trypanosome infection rate in cattle at Nguruman was investigated in a study conducted in 1984-1986. Shifting pastoralism significantly reduced trypanosome infections in cattle. The cattle were more heavily infected with Trypanosoma congolense (16.5%) than Trypanosoma vivax (4.95%) and Trypanosoma brucei (0.19%). Trypanosoma theileri was observed only once among the cattle examined. Mixed trypanosome infections in cattle were observed to be 2.75% and 0.014% for T. congolense/T. vivax and T. congolense/T. brucei, respectively. The duration of infection in the cattle was 55 days for T. congolense and 79 days for T. vivax. High infections in cattle were observed 2 months after the rains, which were concomitant with high tsetse densities.     

Hypothalamic-pituitary-adrenal axis responsiveness to insulin-induced hypoglycaemia is modified by trypanosome infection in Boran (Bos indicus) cattle.

Ten Boran (Bos indicus) cattle were used to study the stress responsiveness of the hypothalamic-pituitary-adrenal (HPA) axis during trypanosome infection. Five cattle were infected with Trypanosoma congolense IL 1180 by tsetse challenge and five cattle served as controls. All infected animals developed acute trypanosomiasis. Insulin-induced hypoglycaemia (50 per cent of pre-insulin glucose concentration) was used as a stress factor. Acute hypoglycaemia was observed in three infected and three control animals after insulin challenge. Two animals from each group either did not respond or responded slowly. Hypoglycaemia in infected animals completely failed to induce an HPA axis response, while in control animals an HPA axis response was indicated by a significant increase in plasma adrenocorticotrophic hormone (ACTH) and cortisol concentrations (P less than 0.01). The results show that trypanosomiasis in Boran cattle can cause a decrease in the stress responsiveness of the HPA axis as indicated by a blunted ACTH/cortisol response to insulin-induced hypoglycaemia.     

Estimation of trypanosomal status by the buffy coat technique and an antibody ELISA for assessment of the impact of trypanosomosis on health and productivity of N'Dama cattle in The Gambia

The buffy coat/dark ground phase contrast technique (BCT) and an indirect antibody enzyme immunoassay (ELISA) were employed to assess the trypanosomal status of 32 N'Dama cattle, aged 19-28 months, exposed to natural challenge of Glossina morsitans submorsitans and G. palpalis gambiensis. Prior to the start of the investigation animals experienced 9-16 months of tsetse challenge in the study area. Blood and corresponding serum samples were examined monthly for a period of 8 months for patent parasitaemia by BCT and presence of Trypanosoma vivax and T. congolense antibodies by ELISA. In the ELISA, the reactivity of sera to anti-trypanosomal antibodies was expressed in percent positivity (pp). Packed cell volumes (PCV) and body weights were also recorded monthly, and daily weight gain (DWG) computed to assess the impact of trypanosomal status on health and productivity. During the study period, the overall parasitaemic trypanosome prevalence was 3% (6/199), while the serological prevalence was 54.7% (109/199). Both diagnostic tests revealed a predominance of T. vivax over T. congolense infections in N'Dama cattle. Sensitivity of the immunoassay was 83.3%. In T. vivax-parasitaemic cattle, antibodies persisted for 4-6 months after the parasite was detected by BCT. A significantly higher overall mean PCV level was observed in blood samples obtained from cattle found, in any particular month, negative by BCT and ELISA, compared with those blood samples from animals responding serologically positively for anti-trypanosome antibodies. Likewise, mean DWG was significantly higher in cattle found negative for both tests in comparison to animals presenting detectable anti-trypanosome antibodies and those detected positive by both tests. A significant negative relationship was observed between pp values and PCV levels in animals seropositive for T. vivax and/or T. congolense. Similarly, a negative relationship was observed between DWGs and pp values. PCV levels were significantly positively correlated with DWGs. It was concluded that serological screening could provide useful information complementary to that obtained by the use of BCT not only to assess more accurately the trypanosomal status of cattle populations, but also to evaluate the effects of trypanosome infection on animal health and productivity and estimate the trypanosomosis risk.     

Trypanosome non-specific IgM antibodies detected in serum of Trypanosoma congolense-infected cattle are polyreactive.

Serum Ig from Trypanosoma congolense-infected cattle were affinity-purified using immobilised trypanosome or non-trypanosome antigens (beta-galactosidase, cytochrome C and ferritin). The bound and unbound IgG and IgM fractions were collected and tested in ELISA for reactivity to each antigen. The results indicated that the presence of reactivity to non-parasite antigens in serum of infected cattle is due to polyreactive IgM antibodies. However, the IgG fraction only bound to trypanosome antigens and was only present in post-infection sera, indicating that it was induced by the infecting trypanosomes. Since the polyreactive IgM antibodies were also present in pre-infection sera, it is probable that they were natural antibodies that were not induced but only amplified by the trypanosome infection.     

Susceptibility and immune responses of zebu and taurine cattle of West Africa to infection with Trypanosoma congolense transmitted by Glossina morsitans centralis

Following tsetse-transmitted infection with Trypanosoma congolense, major differences in development of localised skin reactions, the ability to control parasitaemia, the degree of anaemia and in antibody response to trypanosomes were found between the reputedly trypanotolerant breeds of cattle (N'Dama, N'Dama/Baoule crosses, Baoule) and the trypanosusceptible West African Zebu. The local skin reactions that developed in the Zebu were large and severe while those that occurred in the other breeds were smaller and less severe or mild. The timing of appearance of parasitaemia and the height of the first peaks were similar in all the animals, but the Zebu were less able to control subsequent waves of parasitaemia. Possibly reflecting these events, it was only in the Zebu that significant anaemia developed. Neutralizing antibody against homologous metacyclic trypanosomes developed between 14 to 18 days after infection in all breeds of cattle; however, marked differences were found when antibody to trypanosomes derived from first peak parasitaemias were tested in the Zebu and Baoule. Neutralizing antibody against these parasites appeared in the Baoule on day 24 but were not detected in Zebu until day 51. Furthermore, the antibody titres were 3 log2 higher in the Baoule. It was concluded that the trypanotolerance exhibited by the West African taurine cattle might be related to a) their ability to control trypanosome numbers in the skin and in the bloodstream, an outcome that was possibly brought about by the earlier and superior immune response and b) failure to develop anaemia which might be associated with their capacity to control parasitaemia.     

Evaluation of an indirect fluorescent antibody test, enzyme-linked immunosorbent assay and quantification of immunoglobulins in the diagnosis of bovine trypanosomiasis.

An indirect fluorescent antibody test (IFAT), a microscale version of the enzyme-linked immunosorbent assay (microELISA) and determination of IgM levels in serum were assessed for their comparative diagnostic value in the detection of bovine trypanosomiasis. Serum samples from drug-treated N'dama cattle and untreated N'dama and Zebu cattle from Liberia were examined for the presence fo antibodies to trypanosomes. In the untreated Zebu cattle, infections with T. vivax predominated and the prevalence of infection was higher than that found in untreated N'damas in which infections with T. congolense predominated. The proportion of animals which showed serological evidence of trypanosomiasis in the untreated Zebus was slightly higher than that found in the untreated N'damas. The prevalence of infection was low in N'dama cattle which had been treated with diminazene aceturate and homidium chloride but 50% of the animals showed serological evidence of trypanosomiasis. More serologically positive animals were detected by microELISA than IFAT, but both tests were equally sensitive in detecting antibodies in cattle in which trypanosomes were demonstrated by examination of peripheral blood. With both IFAT and microELISA it was necessary to carry out tests using antigens prepared from T. brucei, T. vivax and T. congolense in order to detect all serologically positive animals. Increases in serum IgM occurred in both N'dama and Zebu cattle but the levels were raised in only approximately half of the known infected animals. Overall, more animals gave positive reactions with IFAT and microELISA than showed raised IgM levels.     

Application of PCR and DNA probes in the characterisation of trypanosomes in the blood of cattle in farms in Morogoro, Tanzania

Polymerase chain reaction (PCR) and deoxyribonucleic acid (DNA) probes were used to characterise trypanosomes from cattle in Morogoro region of Tanzania. Blood samples collected from 390 beef and dairy cattle in selected farms in Morogoro region were examined for presence of trypanosomes using the buffy coat technique (BCT) and blood smears (BSs). Fifty-two animals were found infected: 40 with Trypanosoma congolense, 10 with T. vivax and two with both T. congolense and T. vivax. DNA extracted from all the parasitologically positive and 62 randomly selected parasitologically negative samples were subjected to PCR amplification using primers specific for different trypanosome species. Using a set of seven specific-pairs of primers on the parasitologically positive samples, we detected only T. congolense, either the Savannah- or the Kilifi-type, as single or mixed infections. With the PCR, trypanosome DNA could be detected in 27 (43%) out of 62 samples that were parasitologically negative. DNA hybridisation using probes specific for Savannah- or Kilifi-types T. congolense, or T. vivax, confirmed the presence of these parasites in cattle kept on some farms in Morogoro region of Tanzania. From these studies, it is clear that there is a need to undertake molecular epidemiological studies to determine the distribution of trypanosome species and subspecies, and to assess the economic impact of these parasites in the productivity of livestock in Tanzania. In particular, it would be desirable to verify the assumed association between the different presentations of trypanosomosis on one hand and genotypes of T. congolense on the other.     

Dominance of Chlamydia psittaci-specific IgG2 subclass in the humoral immune responses of naturally and experimentally infected cattle

Indirect enzyme-linked immunosorbent assays were applied to differentiate Chlamydia (C.) psittaci-specific IgG1 and IgG2 levels in 143 individual serum samples from cattle with naturally occurring chlamydial infections and in 190 sequential serum samples from 26 experimentally infected pregnant cows, calves, and a bull. The mean IgG1:IgG2 ratio of naturally infected cattle was 1:4 indicating a significant (p less than 0.001) IgG2 dominance. Similar ratios were detected in the experimentally infected cattle. The dominance of IgG2 was independent of breed, sex, and age. Twenty-nine cattle had significant immunoglobulin levels to both C. psittaci and Coxiella (C.) burnetii simultaneously. The predominance of C. psittaci-specific IgG2, in contrast to the predominance of C. burnetti-specific IgG1 detected in these same individual serum samples under identical conditions, indicates that the ability to preferentially produce either IgG1 or IgG2 was not limited in these individual cattle. A transient yet significant IgG1 response was also developed in cows following chlamydia-induced abortions (immunotype 1) or in cattle infected with the polyarthritis-serositis-encephalomyelitis agents (immunotype 2). IgG1 levels decreased faster than IgG2 levels. These findings have diagnostic implications and identify the need for determining the immunoglobulin classes and subclasses of the humoral immune responses of animals and man to chlamydial infections.     

Cellular and humoral responses in liver of cattle and buffaloes infected with a single dose of Fasciola gigantica

The cellular components of the hepatic inflammatory infiltrate in cattle and buffaloes infected with a single dose of 1000 Fasciola gigantica were analysed by immunohistochemistry and histology. T and B lymphocytes, plasma cells, eosinophils and mast cells were present in the hepatic lesions. It is proposed that both cellular and humoral immune responses were induced in the liver of cattle and buffaloes during infection with F. gigantica probably by antigens released by the developing flukes and by damage caused by the flukes during their migration in the liver. The local T cell response differed between these animals, with the response decreasing after 3 weeks post-infection in cattle in contrast to a gradually increasing response in buffaloes. Difference in the T cell response between cattle and buffaloes may be related to their differences in resistance and resilience to infection with F. gigantica.