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Validation of the FAMACHA eye color chart for detecting clinical anemia in sheep and goats on farms in the southern United States.

Recent studies on sheep and goat farms in the southern United States indicate that multiple-anthelmintic resistance in Haemonchus contortus is becoming a severe problem. Though many factors are involved in the evolution of resistance, the proportion of the parasite population under drug selection is believed to be the single most important factor influencing how rapidly resistance develops. Therefore, where prevention of resistance is an important parallel goal of worm control, it is recommended to leave a portion of the animals untreated. Recently, a novel system called FAMACHA was developed in South Africa, which enables clinical identification of anemic sheep and goats. When H. contortus is the primary parasitic pathogen, this system can be applied on the farm level to reduce the number of treatments administered, thereby increasing the proportion of the worm population in refugia. Since most studies validating the FAMACHA method have been performed in South Africa, it is important that the method be tested in other regions before its use is broadly recommended. We performed a validation study of FAMACHA by testing the system in sheep (n = 847) and goats (n = 537) of various breeds and ages from 39 farms located in Arkansas, Georgia, Louisiana, Florida, and the US Virgin Islands. The color of the ocular conjunctiva of all animals were scored on a 1-5 scale using the FAMACHA card, and blood samples were collected from each animal for determination of packed cell volume (PCV). Fecal samples were also collected from a majority of the animals tested for performance of fecal egg counts (FEC). Correlations between PCV and eye scores, PCV and FEC, and FEC and eye scores were all highly significant for both sheep and goats (P < 0.001). Data for both FAMACHA scores and PCV were evaluated using two separate criteria for anemia: eye score values of 3, 4 and 5 or 4 and 5, and PCV values of < or =19 or < or =15 were considered anemic. Specificity was maximized when eye score values of 4 and 5 were considered anemic and PCV cut off for anemia was < or =19, but sensitivity was low. In contrast, sensitivity was 100% for both sheep and goats when eye score values of 3, 4 and 5 were considered anemic and PCV cut off was < or =15, but specificity was low. In both sheep and goats, predictive value of a negative was greater than 92% for all anemia and eye score categories, and was greater than 99% for both eye score categories when an anemia cutoff of < or =15 was used. Predictive value of a positive test was low under all criteria indicating that many non-anemic animals would be treated using this system. However, compared to conventional dosing practices where all animals are treated, a large proportion of animals would still be left untreated. These data indicate that the FAMACHA method is an extremely useful tool for identifying anemic sheep and goats in the southern US and US Virgin Islands. However, further studies are required to determine optimal strategies for incorporating FAMACHA-based selective treatment protocols into integrated nematode control programs.

Anemia↗

Effect of copper oxide wire particles dosage and feed supplement level on Haemonchus contortus infection in lambs.

The objective of the experiment was to determine the optimal dose of copper oxide wire particles (COWPs) to reduce infection of Haemonchus contortus in male lambs. Five to six-month-old hair breed lambs were housed on concrete and fed 450 (L; n = 25) or 675 g (H; n = 25) corn/soybean meal supplement and bermudagrass hay. In July, lambs were inoculated with 10,000 L(3) larvae (97% H. contortus; Day 0). Lambs were administered 0, 2, 4, or 6 g COWP on Day 28. Concentrations of copper in the liver were determined. There were no effects of supplement level on concentrations of copper in the liver and a linear relationship existed between COWP treatment and concentrations of copper in liver (P < 0.001). Least squares means of the 0, 2, 4, 6 g COWP treatments were 62.2, 135.7, 161.1, and 208.4 ppm (P < 0.001). Between Days 0 and 28, PCV declined and by Day 42, PCV of all COWP-treated lambs was markedly higher than control lambs and remained higher (COWP x day, P < 0.05). By Day 21, PCV was greater in the H compared with the L group of lambs (P < 0.001). Within 14 days of COWP treatment FEC declined from more than 8000 eggs/g (epg) to less than 250 epg in all COWP-treated lambs (COWP x day, P < 0.001). The numbers of H. contortus in the abomasum were greatly reduced in all COWP-treated groups of lambs and remaining nematodes were predominantly males. FEC were greater in L versus H supplemented lambs and values decreased to a greater extent in H lambs when treated with COWP (supplement x COWP; P < 0.02). The 2 g COWP was effective in alleviating H. contortus infection and reducing number of egg-laying nematodes in the abomasum with the lowest concentration of copper in the liver of the COWP treatment groups. PCV values were more favorable for lambs fed the higher level of supplement, especially when FEC were greater than 8000 epg.

Abomasum↗

Evaluation of the phenotypic performance of a Red Maasai and Dorper double backcross resource population: indoor trickle challenge with Haemonchus contortus.

Six F(1) Dorper (D) x Red Maasai (R) rams were mated to both D and R ewes to produce backcross lambs. These six double backcross resource families are being analysed to identify quantitative trait loci that may be controlling resistance to gastro-intestinal (GI) nematode parasites, mainly Haemonchus contortus. After assessing the phenotypic performance of the lambs following exposure to natural infections, the surviving lambs were drenched and moved indoors for an artificial challenge with H. contortus along with straightbred D and R lambs. A total of 1317 lambs were included in the analyses consisting of 523 3/4D, 580 3/4R, 87 D and 127 R. The D lambs were significantly and consistently heavier than R lambs and 3/4D were significantly heavier than the 3/4R lambs. The difference between the backcrosses was about half of that between the straightbreds. Resistance was assessed in terms of faecal egg counts (FEC) and total worm counts (TWC) at necropsy while packed cell volume (PCV) was used to assess resilience to weekly oral doses of 2500 infective larvae of H. contortus. No significant breed differences were observed for log transformed FEC (LFEC). A significant breed difference in PCV was recorded. The backcrosses had the higher values and while no differences were observed between the straightbreds, 3/4D had significantly higher PCV than the 3/4R. Despite the absence of breed differences in FEC the R and the 3/4R had significantly fewer worms than the D and the 3/4D. The D had significantly longer worms than the R and the 3/4D had significantly longer worms than the 3/4R. Worms recovered from D had more eggs than those recovered from R. Similarly worms from 3/4D contained more eggs than those from 3/4R. Thus, on a breed basis the breed with more worms had longer worms. In contrast, when, in a small part of the experiment two doses of larvae were used to check for any breed by dose interactions, worms from the low dose (and hence fewer worms) animals were longer. We postulate that in fast growing hosts like the D, worms also have a better potential for growth and reproduction than in hosts that have less potential for growth. The overall correlation coefficient between PCV and LFEC was -0.67 while that between LFEC and LTWC was 0.72. From these results it is clear that the R do not respond to the artificial challenge the way they do to natural infection suggesting that phenotyping of R and R cross lambs for purposes of selecting those that are resistant or susceptible is best done under natural challenge.

Animals↗

Characterization of moxidectin resistant Trichostrongylus colubriformis and Haemonchus contortus.

The development of moxidectin resistance (MOX-R) in sheep parasitic gastrointestinal nematodes already carrying multiple resistances to other anthelmintic groups has made control of these strains very difficult. The anthelmintic resistance patterns of MOX-R strains of Trichostrongylus colubriformis and Haemonchus contortus were characterized to provide an insight into the remaining role of anthelmintics in the control of such strains. Homozygous MOX-R individuals of both genera were unaffected by moxidectin. For MOX-R heterozygotes a dose rate of 200 microg/kg abamectin (ABA) given orally removed 25% of H. contortus while 200 microg/kg MOX given orally achieved a 72% reduction. Doubling the dose rate of ABA improved the mean efficacy to 37%. Consequently, in H. contortus, the degree of dominance differs markedly between the two anthelmintics. A dose rate of 8 mg/kg levamisole and 185 mg/kg napthalophos achieved >95% reduction in worm count of the MOX-R homozygous H. contortus but only 85 and 7%, respectively against the MOX-R homozygous T. colubriformis.

Abomasum↗

The modulatory influence of Trypanosoma brucei on challenge infection with Haemonchus contortus in Nigerian West African Dwarf goats segregated into weak and strong responders to the nematode.

Although Nigerian West African Dwarf (WAD) goats are relatively resistant to infection with Haemonchus contortus and are also trypanotolerant, natural outbreaks of both infections are known to occur. Despite their relative resistance to H. contortus WAD goats nevertheless show variability in response phenotype and it was of interest to examine the effect of this variability on the outcome of concurrent trypanosome infection. Trypanosoma brucei infections were established in goats that were initially classified as good or poor responders to H. contortus. Thirty-nine goats were exposed to an escalating infection with H. contortus, and on the basis of their mean faecal egg counts (FEC) were allocated to high FEC (poor responders, 18 goats with the highest FEC) or low FEC (good responders, 18 goats with the lowest FEC) classes. Nine uninfected naive control goats were included to provide reference baseline values. Retrospective analysis of parasitological and pathological parameters after allocation into high/low FEC classes showed that FECs differed significantly, in both classes packed cell volume (PCV) values fell relative to naive controls, neither class lost weight and both generated marked IgG responses. All goats received anthelmintic on day 61, half of each group was infected with 50 million trypanosomes and on day 67, excepting the controls, all goats were challenged with 3000 L3 of H. contortus. Trypanosome parasitaemia was generally low, and marginally, but not significantly, higher in the low compared with high FEC class, peaking 12-16 days after exposure in both groups and then falling to below microscopically detectable levels (although still detectable by sub-inoculation into mice) by week 3. At autopsy (days 109/110), worm burdens were significantly higher in the trypanosome-infected goats from the high FEC class, relative to all other groups. Trypanosome infected goats showed a tendency (although not significant) towards higher FEC and, irrespective of their FEC class, had lower PCV values although body weight did not vary significantly. All goats challenged with H. contortus had higher antibody levels than naive controls, but neither trypanosome infection nor FEC class affected the magnitude of responses. These results confirm that WAD goats comprise a range of response phenotypes to initial H. contortus infection and that trypanotolerance is a key trait of this breed. Although immunity to nematode infection develops even in poor responders, these animals harbour higher nematode burdens during concurrent infection with T. brucei.

Animals↗

Relationship of abomasal histology and parasite-specific immunoglobulin A with the resistance to Haemonchus contortus infection in three breeds of sheep.

This study was carried out to evaluate the relationship of abomasal inflammatory cells and parasite-specific immunoglobulin A (IgA) in mucus, with the resistance to Haemonchus contortus infection in three breeds of sheep naturally infected with gastrointestinal nematodes. The breeds were the native Santa Ines sheep, and the European Suffolk and Ile de France breeds. Mast cells, eosinophils and globule leucocytes were enumerated in abomasal mucosa. Eosinophils within the sub-mucosa also were counted separately. Histamine concentration was estimated in abomasal tissue samples. Enzyme-linked immunosorbent assay was carried out in mucus samples to determine the level of IgA anti-H. contortus third and fifth instar. There were no significant differences among group means of these variables (P>0.05). The correlation coefficients between fecal egg counts (FEC)xmast cells (r=-0.490; P<0.05) and FECxeosinophils in sub-mucosa (r=-0.714; P<0.01) was significant in the Santa Ines sheep. In the Ile de France group, the correlation coefficients between globule leucocytesxFEC (r=-0.879; P<0.001) and histaminexworm burden (r=-0.833; P<0.01) were also significant. In the Santa Ines and Ile de France sheep, correlation coefficients between IgA anti-L3xworm burden and IgA anti-L3xFEC were negative. In general, inflammatory cells and IgA-parasite-specific in abomasum were inversely associated with H. contortus worm burden and FEC indicating that they may impair parasite development or fecundity in the three breeds of sheep. However, similar mean values of inflammatory cells and IgA were found in the resistant (Santa Ines) and in the susceptible (Suffolk and Ile de France) breeds of sheep. The enumeration of cells by histological assessment does not provide information on their functional activity, which may be different among breeds. Thus, the effect of breed on the functional activity of these and other inflammatory cells is an important area for further study.

Abomasum↗

Haemonchus contortus egg excretion and female length reduction in sheep previously infected with Oestrus ovis (Diptera: Oestridae) larvae.

Mixed parasitic infection of animals is a common phenomenon in nature. The existence of one species often positively or negatively influences the survival of the other. Our experimental study was started with the objectives to demonstrate the interaction of Haemonchus contortus and Oestrus ovis in relation to cellular and humoral immune responses in sheep. Twenty-two sheep of Tarasconnais breed (France) were divided into four groups (O, OH, H and C) of five or six animals. Group O and OH received 5 weekly consecutive inoculations with O. ovis L1 larvae (total = 82 L1) in the first phase of the experiment between days 0 and 28. On the second phase, groups OH and H received 5000 L3 of H. contortus on day 48 while group C served as our control throughout the experimental period. Parasitological, haematological, serological and histopathological examinations were made according to standard procedures and all animals were slaughtered at day 95. There was no significant variation in the number and degree of development of O. ovis larvae between the two infected groups. Furthermore, in tissues examined in the upper respiratory tract (nasal septum, turbinate, ethmoide and sinus), group O and OH has responded similarly on the basis of cellular inflammatory responses (blood and tissue eosinophils, mast cells and globule leucocytes (GL)) and serum antibody responses against the nasal bots. This may indicate that the presence of H. contortus in the abomasa of group OH had no marked influence over the development of O. ovis larvae in the upper respiratory tract. On the other hand, we have observed a significantly lower H. contortus female worm length, fecal egg count (FEC) and in utero egg count in animals harbouring the nasal bot (OH) than in the mono-infected group (H). This was significantly associated with higher blood eosinophilia, higher packed cell volume (PCV) and increased number of tissue eosinophils and globule leucocytes. We conclude that, the establishment of O. ovis larvae in the upper respiratory tract has initiated higher inflammatory cellular activity in group OH there by influencing the development and fecundity of H. contortus in the abomasum.

Animals↗

Effect of ivermectin on feeding by Haemonchus contortus in vivo.

While several in vitro studies have shown that the anthelmintic ivermectin inhibits feeding by parasites, the relevance of this putative site of action in vivo has not been demonstrated. For this study, techniques to measure feeding by Haemonchus contortus in vivo relied on the blood feeding characteristics of the worm, and utilised tritiated inulin administered to sheep intravenously and subsequently measured in worms recovered from abomasa. Nematodes recovered from sheep treated with ivermectin 4 h prior to the [3H]inulin administration showed equivalent feeding levels (over a 1 h period) to those recovered from sheep not treated with ivermectin. In addition, there was no difference in the radioactivity in nematodes of an ivermectin-susceptible and an ivermectin-resistant isolate recovered from individual sheep with concurrent infections after a dose with ivermectin. Ivermectin, therefore, had no effect on feeding by H. contortus in vivo under these experimental conditions. The results are discussed in relation to the dynamics of the expulsion of H. contortus from sheep following ivermectin treatment.

Abomasum↗

Detection of anthelmintic resistance: a comparison of mathematical techniques.

Anthelmintic resistance has become an increasing problem particularly to gastrointestinal tract nematodes and appropriate methods are required to detect this phenomenon so the correct action can be taken. This paper compares a number of mathematical techniques that are used to analyse data. The negative binomial distribution is a mathematical distribution used to model aggregated data and hence is suitable to model the intensity of parasite burden and the magnitude of the faecal egg counts. Maximum likelihood techniques are utilised to exploit this mathematical distribution to analyse the magnitude of the faecal egg count reduction and decline in the worm burden in response to anthelmintic treatment. Data from experimental groups of sheep described in the accompanying paper are used. In addition, simulated data sets of faecal egg counts were created using a random number generator following appropriate negative binomial distributions. The results demonstrate this statistical model can detect evidence of anthelmintic resistance with a faecal egg reduction test that otherwise might require a slaughter trial to demonstrate. In addition, the simulated data sets confirm that there is a significant probability of failure to detect low anthelmintic efficacy with commonly used mathematical techniques. Consequently, the use of maximum likelihood mathematical techniques with a negative binomial statistical model would aid in the early detection of anthelmintic resistance using faecal egg count reductions and result in a lower probability of inappropriately assigning an anthelmintic as effective.

Animals↗

Multiple anthelmintic resistance in Haemonchus contortus isolated from South African Boer goats in Switzerland.

A suspected case of multiple anthelmintic resistance on a farm in the canton of Zurich, Switzerland, into which South African Boer goats had previously been imported, was confirmed in a controlled test. Twenty sheep were allocated into one control group and three treatment groups to determine the efficacy of mebendazole, ivermectin and moxidectin applying the faecal egg count reduction test (FECRT). The sheep were slaughtered 1 week later and post-mortem worm counts were performed. Benzimidazole and ivermectin resistance were found in Haemonchus contortus with an efficacy of 55 and 61%, respectively. Moxidectin appeared to be effective when the data was analysed according to the recommended analytical techniques with an efficacy of 96%. This is the first description of resistance of gastrointestinal nematodes to one of the macrocyclic lactones in small ruminants in Switzerland. The results are discussed in relation to the importance of controlling livestock before importation.

Abomasum↗

Rapid selection for ivermectin resistance in Haemonchus contortus.

Lambs infected with two isolates, one British and one American, of Haemonchus contortus were treated with increasing doses of ivermectin. Eggs from the highest dose that had not eliminated the infection were cultured and larvae used to infect another lamb. After three generations the H. contortus was resistant to 0.2 mg/kg ivermectin. The results stress the ease with which ivermectin resistance can be selected if high selection pressure is applied.

Animals↗

The predatory capability of Arthrobotrys cladodes var. macroides in the control of Haemonchus contortus infective larvae.

One hundred compost samples were examined for the presence of nematophagous fungi on the sheep farms of Mazanderan, province, Iran. Arthrobotrys cladodes var. macroides (IRAN 677C=CBS 143565) was isolated from 3% of the samples examined. Nematophagous activity of this fungus which was shown for the first time in this study, revealed the addition of 1000, 8000, 20000 and 100000 conidia per gram of feces of sheep reduced significantly (P<0.001) the number of Haemonchus contortus infective larvae in the feces by 41.71%, 63, 27%, 73.49% and 94.96%, respectively. These results show that A. cladodes var. macroides is a promising candidate for biological control of H. contortus.

Animals↗

Consequences of the unusually warm and dry summer of 2003 in The Netherlands: poor development of free living stages, normal survival of infective larvae and long survival of adult gastrointestinal nematodes of sheep.

During 2003 a grazing study was performed at Utrecht University to evaluate evasive grazing and application of Duddingtonia flagrans for the control of parasitic gastroenteritis in sheep. However, the summer of 2003 (June-August) was unusually warm and dry. As a result the patterns of gastrointestinal nematode infections deviated from those observed in more average years. The proportion of eggs that developed to infective larvae was far lower than normal in July-August. On the other hand, survival of larvae that had developed before the middle of July was not affected compared to other years. In fact, severe haemonchosis was observed in (tracer) lambs grazed at the end of July on pastures that had been contaminated from 26-05 to 16-06 and from 16-06 to 07-07. Moreover, tracer lambs grazing in September on some of these plots still acquired large Haemonchus contortus burdens. Over 60% of H. contortus that had established before the middle of July appeared to be able to survive until October in the virtual absence of re-infection. That may have consequences for the application of evasive grazing as a control option in that suppression of adult burdens might still be necessary, through a limited use of anthelmintics or through alternative deworming strategies.

Animal Husbandry↗

Influence of dietary protein supply on resistance to experimental infections with Haemonchus contortus in Ile de France and Santa Ines lambs.

The effect of Haemonchus contortus infection in sheep fed with a moderate and high protein content diet was evaluated in two breeds of sheep. Forty-eight Ile de France and Santa Ines lambs were maintained indoors since birth, in worm-free conditions. The lambs were allocated after weaning in four groups of six animals per breed, which were either infected or remain uninfected and given access to either a moderately or highly metabolizable protein diet. The moderately and highly metabolizable protein diets were calculated to supply 75 and 129 g metabolizable protein per kg of dry matter (MP/kg DM), respectively. The infection consisted of a trickle infection with 300 infective larvae, three times a week, for 12 weeks. Significant differences were observed for mast cell, globule leukocyte and eosinophil counts in the abomasal mucosa of the infected groups compared to the control of both breeds (P<0.05), regardless of the diet supplied. Significantly higher IgA anti-L5 antibody was detected in the infected Santa Ines groups than in the infected Ile de France groups (P<0.05). Increased metabolizable protein supply resulted in larger body weight gain and higher packed cell volumes for both breeds (P<0.05). Both breeds showed an increased ability to withstand the pathophysiological effects of H. contortus infection when given access to the highly metabolizable protein diet. However, increased metabolizable protein supply resulted in reduced worm burdens in Santa Ines lambs but not in the Ile de France lambs (P<0.05). The present results show that the increase in protein content in growing lamb diets may benefit resistance and resilience to gastrointestinal parasites but that these benefits may vary among breeds.

Abomasum↗

Interaction between copper oxide wire particles and Duddingtonia flagrans in lambs.

An experiment was completed to determine if copper oxide wire particles (COWP) had any effect on the activity of the nematode-trapping fungus Duddingtonia flagrans in growing lambs. COWP has been used recently as a dewormer in small ruminants because of nematode resistance to anthelmintics. D. flagrans has been used to control free-living stages of parasitic nematodes in livestock. Katahdin and Dorper lambs, 4 months of age, were administered no or 4 g COWP (n=24/dose) in early October 2003. Haemonchus contortus was the predominant gastrointestinal parasite during the trial, which was acquired naturally from pasture. Half the lambs from each COWP group were supplemented with corn/soybean meal with or without D. flagrans for 35 days. Fecal egg counts (FEC) and packed cell volume (PCV) were determined weekly between days 0 (day of COWP administration) and 35. Feces from lambs in each treatment group were pooled and three replicates per group were cultured for 14 days at room temperature. Larvae (L3) were identified and counted per gram of feces cultured. Treatment with COWP was effective in decreasing FEC, which remained low compared with FEC from lambs not treated with COWP. This led to an increase in PCV in these lambs (COWP x day, P<0.001). Number of larvae was decreased in feces from lambs treated with COWP and D. flagrans between days 14 and 35 compared to the other groups of lambs (COWP x D. flagrans x day, P<0.003). Percentage of larvae identified as H. contortus decreased in feces collected from lambs treated with COWP and D. flagrans between days 14 and 28 compared with other treatments (COWP x D. flagrans x day, P<0.05). Other trichostrongyles were present and remained less than 7% in feces collected from control lambs. There was no adverse effect of COWP on the ability of D. flagrans to trap residual larvae after COWP treatment. With fewer eggs being excreted due to the effect of copper on H. contortus, and the additional larval reducing effect exerted by the nematode destroying fungus D. flagrans, the expected result would be a much lower larval challenge on pasture when these two tools are used together in a sustainable control strategy.

Animals↗

Haemonchus contortus and Trichostrongylus colubriformis in pen-trials with Javanese thin tail sheep and Kacang cross Etawah goats.

Weight gain costs due to infection were higher in sheep than goats, 28 and 17.5%, respectively, for Trichostrongylus colubriformis and 48.7 and 32.2%, respectively, for Haemonchus contortus. The extent of bodyweight cost attributed to anorexia in sheep infected with H. contortus was higher (13.5 g/day) than in sheep infected with T. colubriformis (2.3 g/day). On the other hand, bodyweight cost due to the other pathogenic effects in sheep infected with T. colubriformis were higher (35.6 g/day) compared to sheep infected with H. contortus (10.9 g/day). A strong relationship between faecal egg count and worm count (r=0.79, P=0.006) was shown only in sheep infected with T. colubriformis. About half of the infected sheep and goats had low or zero faecal egg counts throughout the study. In about 40% the egg count rose initially but became low by weeks 10-16, whereas in about 10% counts increased progressively throughout the period of observation and these animals also had the highest numbers of worms at slaughter. Packed cell volume was reduced in sheep and goats infected with H. contortus but serum protein and haemoglobin levels were unaffected. Sheep infected with T. colubriformis had a higher level of eosinophilia after 8 weeks (18.4%) than sheep infected with H. contortus (11.4%), whereas this pattern was reversed in goats and levels were also lower (4.1 and 8.9%, respectively). There was no apparent relationship between eosinophilia and resistance to infection with H. contortus or T. colubriformis.

Animals↗

Towards the eradication of Haemonchus contortus from sheep flocks in Sweden.

Previous studies have shown that between-year transmission of Haemonchus contortus in Swedish sheep flocks is almost entirely as over-wintered populations within housed animals, and not on pasture. This suggests that eradication of this parasite is a realistic possibility. Thus, two sheep farms with a history of H. contortus infection on the Swedish island of Oland were selected for study. During the winter housing period of 2003/2004 all ruminants (sheep and cattle) on both farms were treated with ivermectin. Monitoring by faecal egg counts and infective larval differentials of ewes and lambs for the subsequent two grazing seasons, together with total abomasal worm counts of 10 lambs from each farm at the end of the first grazing year, showed that this objective was achieved.

Animals↗

Effect of CD4+ T lymphocyte depletion on resistance of Gulf Coast Native lambs to Haemonchus contortus infection.

It has been reported that CD4(+) T lymphocytes are important in acquired immunity to gastrointestinal nematode infection. Whether these lymphocytes are also involved in the immune response of naturally resistant Gulf Coast Native (GCN) sheep to Haemonchus contortus infection remains to be defined. The objective of this study was to determine the role of CD4(+) T lymphocytes in this resistance. Ten GCN lambs were randomly assigned to a control (n=5) or a treatment (n=5) group. The treatment consisted of a series of IV injections with mouse anti-ovine CD4(+) T lymphocyte monoclonal antibodies for a period of 3 weeks. After the second treatment, all lambs were experimentally infected with 10,000 H. contortus infective larvae by oral inoculation. All lambs were monitored for fecal egg counts, blood packed cell volumes, white blood cell differential counts and serum antibody responses on a weekly basis. Fluorescence-activated cell sorter (FACS) analysis was done biweekly to enumerate CD4(+) T lymphocytes in peripheral blood. Necropsies were performed at the end of the study and 10% of the contents of the gastrointestinal tract were preserved for nematode enumeration and identification. Also at necropsy, mesenteric lymph nodes were extracted and FACS analysis was run on lymphoid cells. Mean fecal egg counts on day 21 and 28 post-infection and nematode counts at necropsy of the treated group were significantly (p<0.05) higher than that of the control group. Percent CD4(+) T lymphocytes in peripheral blood was significantly (p<0.05) lower in the treatment group than in the control group from day 9 to the end of the study. No differences were found in blood packed cell volumes, white blood cell differential counts, antibody titer or lymph node CD4(+) lymphocytes between groups. Lambs depleted of their CD4(+) T lymphocytes were more susceptible to H. contortus infection than undepleted lambs. The results of this study suggest that CD4(+) T lymphocytes are associated with the natural resistance of GCN sheep to H. contortus infection.

Abomasum↗