捻转血矛线虫重组H11抗原山羊免疫保护性试验

以重组H11-1、H11-2及H11-1与H11-2混合物分别免疫山羊后,ELISA检测发现血清抗体均在首次免疫后14 d达到较高水平,在第2次免疫后10 d达到高峰。用10 000只捻转血矛线虫第3期幼虫攻击山羊后,对不同免疫组山羊排出虫卵数、虫卵孵化率和成虫数的统计分析结果表明,免疫组与对照组间差异不显著,但免疫组雌虫数显著少于对照组,这对防治捻转血矛线虫病可能会起一定的作用。     

Variability of resistance in goats infected with Haemonchus contortus in Brazil

The variability between and within breeds with respect to nematode egg counts (EPG), packed cell volume (PCV) and hemoglobin (Hb) was studied in 36 yearling female goats of the Caninde (15), Bhuj (6) and Anglo-Nubian (15) breeds, exposed to Haemonchus contortus. Nematode-free goats were turned to a contaminated paddock in late February. From then on, fecal egg per gram counts (EPG), packed cell volume (PCV) and hemoglobin (Hb) were determined at 2-week intervals up to Week 18. The EPG, transformed as [log(EPG + 75)], varied (P < 0.01) between goats within breeds and between weeks of exposure, but not between goat breeds (P > 0.05). PCV and Hb were affected by goat breeds (P < 0.05), by goats within breeds (P < 0.01) and by weeks of exposure (P < 0.01). Anglo-Nubians had higher (P < 0.01) PCV and Hb than Caninde; Bhuj had intermediate values. There were two EPG rises; one between Weeks 6 and 10 and the other between Weeks 14 and 16. The within breed variability was marked during the EPG rise on Week 6, when individual egg counts ranged from 130 to 2500. The EPG rises coincided with drops in Hb. PCV presented a similar trend, though not as marked. Haemonchus was responsible for more than 95% of nematode eggs counted. Considering the goat as experimental unit, the correlation coefficients (r) were: -0.45, P = 0.0064, between log(EPG + 75) and PCV; and -0.53, P = 0.0009, between log(EPG + 75) and Hb. The negative correlation between egg counts and blood values suggested breed differences in PCV and Hb were related to resistance to H. contortus infection and/or to its effects.     

The establishment and development of Haemonchus contortus in goats

Twelve goats were inoculated with 40,000 third-stage Haemonchus contortus larvae and two were killed on each of Days 4, 7, 11, 14, 18 and 21 after inoculation (DAI). The number of worms that established, and the site of development were recorded. More worms established in the fundic, than in the middle or pyloric thirds of the abomasum. Early development occurred within the mucosa; emergence into the lumen started between 7 and 11 days after infection. By 4 DAI, all worms had completed the third moult to the L4 stage. At 11 DAI the majority of the worms were adults. A mean of 13.2% of the female worms had eggs in their uteri at 18 DAI; by 21 DAI more than half of the female worms had eggs in their uteri. The development of H. contortus was essentially similar to that described in sheep.     

Effects of condensed tannins on goats experimentally infected with Haemonchus contortus

Although the use of tanniferous plants or condensed tannins as an alternative to anthelmintics to control gastrointestinal nematodes has been largely documented in sheep, studies remain scarce in goats. The objective of this study was therefore to assess the possible impact of condensed tannins in goats infected with adult Haemonchus contortus. Two groups of cull goats were experimentally infected with 10.000 L3 of H. contortus. After 4 weeks, quebracho extracts, representing 5% of the diet DM, were administered for 8 days to one of the two groups. Goats of the second group remained as controls. One week after the end of quebracho administration, the goats were euthanised. Individual egg excretion and pathophysiological parameters were measured weekly during the study. At the end of the study, worm counts were assessed and histological samples from the abomasa were taken to count the numbers of mucosal mast cells, globule leukocytes and eosinophils. The administration of tannins was associated with a significant decrease in egg excretion, which persisted until the end of experiment. This reduction was not associated with any difference in worm number but with a significant decrease in female fecundity. No significant changes in the mucosal density of the three inflammatory cell types were detected between the two groups. These results indicate that the major consequence of tannin consumption in goats is a reduction in worm fecundity and egg output, which does not seem related to significant changes in the local mucosal response.     

Effect of feeding sericea lespedeza leaf meal in goats experimentally infected with Haemonchus contortus

Effect of sericea lespedeza [SL; Lespedeza cuneata (Dum-Cours.) G. Don.] leaf meal feeding was evaluated in two experiments in indoor reared goats with experimental infection of Haemonchus contortus larvae. In the first experiment, ten 8-10 month old male Spanish and Alpine cross kids pair matched for body weight and age were fed SL or bermudagrass [BG; Cynodon dactylon (L.) Pers.] hay one week before infection and were infected with 5000 H. contortus L(3). The animals were maintained on the same diet for the remaining period and were slaughtered 28 days post-infection (DPI) to determine the establishment of incoming infective larvae. Goats fed SL had lower establishment (P<0.05) of H. contortus larvae than that of the control goats fed BG hay. In the second experiment, twenty-five 8-10 months old male Alpine cross, Saanen, Nubian×Saanen and Spanish kids reared in confinement on BG were experimentally infected with 5000 H. contortus L(3). On 35 DPI, the animals were allocated to two groups after blocking by fecal egg count (FEC), and one group was fed SL leaf meal (n=13), and another control group remained on BG (n=12). Four goats/group were slaughtered successively on days 7, 14, and 28 days post SL feeding, except on day 7, when five SL fed goats were slaughtered. Fecal egg counts and blood packed cell volume (PCV) were measured at weekly intervals and worm count, female worm fecundity, worm length and mucosal eosinophils, mast cells and globule leucocytes were measured after slaughter. Goats fed SL had a lower FEC (P<0.05) one week after feeding, as compared to those fed on BG, and the values remained at low level thereafter. Similarly, PCV was also significantly affected by feeding (P<0.01), and feeding and time interaction (P<0.05). However, worm burden, female worm fecundity, parasite length, and mucosal inflammatory cell count were similar between the groups. Feeding SL reduced the establishment of infective larvae and FEC of H. contortus in experimental studies and this plant could be used for biological control of parasite infection under field conditions to limit the harmful effects of the parasites in goats.     

绵阳地区山羊粪便中捻转血矛线虫的分子鉴定

为鉴定绵阳地区山羊粪便中疑似血矛属线虫虫卵的虫种,本研究提取绵阳地区三个县山羊粪便中疑似血矛属线虫的虫卵DNA 60份,采用PCR技术扩增其核糖体第二内转录间隔区(rDNA ITS-2)基因序列,并与GenBank上公布的捻转血矛线虫同源序列进行比较分析。结果显示:60份疑似血矛属线虫的虫卵DNA样品均得到191bp的ITS-2片段;60条ITS-2序列共形成15条不同的序列,含有17个单变异位点,与GenBank中多株捻转血矛线虫的同源性达95.8%~100%;ML系统发育树显示,本研究得到的15种类型ITS-2序列与已报道的捻转血矛线虫(Haemonchus contortus)聚类形成一个分支。综上可得,60份来自山羊的血矛属线虫均为捻转血矛线虫。     

In vivo anthelmintic activity of Phytolacca icosandra against Haemonchus contortus in goats

Two separate controlled and blinded studies were conducted to confirm the dose and non-interference of spinosad and milbemycin oxime (MO) administered orally in combination or alone to dogs for the treatment and control of experimentally induced flea infestations (Ctenocephalides felis) and adult hookworm infections (Ancylostoma caninum). For each study, dogs were allocated randomly based on pre-treatment adult flea and hookworm egg counts to one of four treatment groups of 10 animals each. In each study, spinosad and MO in combination, using the lower half (30-45 mg/kg spinosad; 0.5-0.75 mg/kg MO) of the US commercial dose band (30-60 mg/kg spinosad; 0.5-1.0mg/kg MO) of each active ingredient, or individually alone using the full dose range, were given orally to dogs on Day 0 using a tablet formulation. A placebo control was treated similarly. In one study, on Days -1, 5, 12, 19, 28 and 35 each dog was infested with approximately 100 unfed adult C. felis obtained from the investigator's established flea colony. All dogs were infested via the same method. Forty-eight hour post-infestation flea comb counts were conducted on Days 1, 7, 14, 21, 30 and 37 and were used to determine the knockdown and residual flea activity. In the second study, on Day -27 each of 48 dogs were experimentally inoculated with 100 third-stage infective larvae of the hookworm, A. caninum. Dogs were treated on Day 0 and necropsied on Day 7 or Day 8. All nematodes in the intestinal tract were collected on Day 7 or Day 8, identified and counted by species and stage. Post-treatment, the geometric mean live flea counts were significantly different (p-value<0.0001) between the spinosad/MO combination and the spinosad only treatment groups as compared to the vehicle control group. The flea counts in the MO only group and the control group were not statistically different. The spinosad and MO combination group and the spinosad only treatment group demonstrated significantly different knockdown (100%) and post-treatment residual flea efficacy at Day 30 was 100% for both groups as compared to the vehicle control. The presence of MO in combination with spinosad did not interfere with the flea efficacy of spinosad as compared to the spinosad only group. MO alone did not demonstrate any flea efficacy. Post-treatment, the geometric mean A. caninum worm counts were significantly different (p-value<0.0001) between the spinosad and MO combination group as compared to the vehicle control group. The worm counts in the MO only group and the combination group were not statistically different. The spinosad and MO combination group (99.8% reduction) and the MO only treatment group (99.5% reduction) both demonstrated significantly different hookworm efficacy as compared to the vehicle control group. The presence of spinosad in combination with MO did not interfere with the hookworm efficacy of MO as compared to the MO only group. Spinosad alone did not demonstrate any hookworm efficacy. In summary, flavored spinosad and MO combination tablets administered orally to dogs at the lower end (30-45 mg/kg spinosad; 0.5-0.75 mg/kg MO) of the US commercial tablet unit dose range (30-60 mg/kg spinosad; 0.5-1.0mg/kg MO) were both safe and highly efficacious delivering 100% knockdown and 30 days of residual adult flea control on experimentally infested dogs as well as >99% adult hookworm efficacy evaluated under laboratory conditions. Interference between either drugs was not demonstrated for both of these dose limiting parasites.     

Effects of Acacia nilotica and Acacia karoo diets on Haemonchus contortus infection in goats

This study was carried out to determine the effects of Acacia karoo and Acacia nilotica diets on Haemonchus contortus infections in goats. Twenty-four Boer goats of mixed sex (live weight 17-22 kg) were randomly assigned to four treatment groups, namely: A. nilotica (AN) group, A. karoo (AK) group, control infected with H. contortus (HC) group and the non-infected control (NHC) group. Animals in the AN, AK and HC groups were orally infected with a single dose of 3000 HC third stage larvae. The AN and AK groups had dried leaves of AN and AK, respectively, included in their basal diet at a rate of 40% dry matter (DM) while the HC and NHC groups had the basal diet throughout the experiment. All animals received a total feed allowance of 500 g DM per day and Katambora Rhodes grass hay ad libitum for roughage. Parameters measured included faecal egg counts and live weight. At the end of the experiment, three animals from each group were slaughtered and abomasal worm burdens were determined. A significant decrease in the faecal egg counts was recorded in animals in the AK group (P<0.05) relative to those in the AN and HC groups. The worm burdens were reduced by 34% in the AK group (P<0.05) and by 10% in the AN group (non-significant) relative to the infected control group. The study indicates that the difference in the effects of the two forages on HC infections may be related to type and concentration of tannins.     

Single nucleotide polymorphisms in candidate genes are significantly associated with resistance to Haemonchus contortus infection in goats

Background: Haemonchosis is a major economic problem in goat production in humid, tropical and subtropical regions. The disease is caused by an abomasal nematode, Haemonchus contortus, which is highly pathogenic in small ruminants. The aim of this study was to identifying single-nucleotide polymorphisms(SNP) that were associated with fecal egg counts(FEC) and could be used as markers to identify resistance to H. contortus in goats.Results: Ten novel variants in the CI TA, ATP2 A3, HSPA8, STAT5 B, ESYT1, and SERPING1 genes were associated with FEC in goats with a nominal significance level of P 0.05. Two missense mutation in the exon region of the caprine CI TA gene resulted in replacement of arginine with cysteine at position 9473550(R9473550 C) and aspartic acid with glutamic acid at position 9473870(D9473870 E). Chinese goat breeds had significantly higher FEC than Bangladeshi goat breeds within their respective genotypes. Polymorphism information content(PIC), effective al ele number(Ne), and heterozygosity(He)were greatest for the STAT5 B_197_A G SNP locus in al goat breeds. Pairwise coefficients of linkage disequilibrium(D′, r~2)revealed complete LD(r~2= 1) between significant SNP polymorphisms in CI TA and SERPING1 and strong LD(r~2= 0.93 and0.98) between polymorphisms in HSPA8 and ATP2 A3, respectively. Correlation coefficient(r) between FEC and body weight(BW) was significantly positive(r = 0.56***, P 0.001) but that between FEC and packed cel volume(PCV) was negatively significant(r =-0.47**, P 0.01) in the total population of goats. On the other hand, correlation coefficient(r)between BW and PCV was not significant in total population of goats. Association analysis revealed that haplotypes within ATP2 A3, HSPA8, and SERPING1 were significantly associated with FEC. Quantitative real-time PCR revealed that the relative expression of m RNA was higher(P 0.001) for resistant, compared to susceptible, groups of goats for al candidate genes except CI TA.Conclusions: This study identified SNP markers that can potential y be used in marker-assisted selection programs to develop goat breeds that are resistant to H. contortus.     

Evaluation of ivermectin against experimental infections of Haemonchus contortus and Trichostrongylus colubriformis in goats.

Fourteen nematode parasite-na?ve kids were given a mixed larval inoculum (approximately 17,250 infective larvae) via stomach tube. Twenty-two days after infection, the kids were randomly assigned to control and treatment groups of seven animals each. Mean eggs per gram for control and treatment groups were 4304 and 5315, respectively. Kids in the treatment group were given ivermectin (0.2 mg kg-1) administered by subcutaneous injection. At necropsy, the mean numbers of Haemonchus contortus and Trichostrongylus colubriformis for control and treatment groups were 2259 and 0, and 3033 and 773, respectively. This reduction was significant (P less than 0.05) for both species, resulting in an efficacy of 100.0% and 74.5%, respectively. The lower efficacy against T. colubriformis may be a sign of resistance, a reduced effectiveness due to route of administration, or a higher dose may be required with subcutaneous administration, as has been observed previously.     

Evaluation of copper supplementation to control Haemonchus contortus infections of sheep in Sweden

A pen study was conducted to assess the effect of providing daily copper mineral supplement, or copper wire particle (COWP) capsules, on established or incoming mixed nematode infections in young sheep. For lambs with established (6 week old) infections, COWP resulted in 97% and 56% reduction of the adult and early L4 stages of H. contortus, respectively, compared with controls (p<0.001). Additionally there was a 74% reduction in Teladorsagia circumcincta infections in the COWP lambs compared with controls (p<0.01). However, no effect was observed when COWP were given at the commencement of a larval dosing period of 6 weeks. There was no significant effect of copper mineral supplement (given at the recommended rate to prevent Cu deficiency) on either established, or developing parasite infections. In addition, a field trial was conducted on a commercial farm to assess the effects of COWP in the management of recurrent H. contortus infections, but lack of parasites during the grazing season prevented an adequate assessment from being made. These results indicate that there is little, if any, benefit from a parasite control standpoint in recommending copper therapy, specifically to control parasites in Swedish sheep flocks.     

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.     

Experiences with integrated concepts for the control of Haemonchus contortus in sheep and goats in the United States

The generally warm, moist environmental conditions in the southern United States (U.S.) are ideal for survival and growth of the egg and larval stages of Haemonchus contortus and other gastrointestinal nematodes (GIN) of sheep and goats. Consequently, infection with GIN is the greatest threat to economic small ruminant production in this region. With anthelmintic resistance now reaching epidemic proportions in small ruminants in the U.S., non-chemical control alternatives are critically needed. The Southern Consortium for Small Ruminant Parasite Control (SCSRPC) was formed in response to this crisis and over the last decade has successfully validated the use of several novel control technologies, including FAMACHA(?) for the implementation of targeted selective treatments (TST), copper oxide wire particles (COWP), nematode-trapping fungi, and grazing or feeding hay of the high-tannin perennial legume sericea lespedeza [Lespedeza cuneata (Dum.-Cours. G. Don)]. Producer attitudes toward GIN control in the U.S. have been shifting away from exclusive dependence upon anthelmintics toward more sustainable, integrated systems of parasite control. Some novel control technologies have been readily adopted by producers in combination with appropriate diagnostic tools, such as FAMACHA(?). Others techniques are still being developed, and will be available for producer use as they are validated. Although new drugs will likely be available to U.S. goat and sheep producers in the future, these will also be subject to development of anthelmintic resistance. Therefore, the adoption and implementation of sustainable GIN control principles will remain important. With emerging markets for grass-fed or organic livestock, there will continue to be a critical need for research and outreach on development and on-farm application of integrated GIN control systems for small ruminants in the U.S. for the foreseeable future.     

Changes in liveweight gain and blood constituents in experimental infection of goats with a goat-derived compared with a sheep-derived strain of Haemonchus contortus.

Two groups of goats were experimentally infected with two different strains of Haemonchus contortus and compared with the controls. Group A animals were infected with a goat-derived strain (GDS) while Group B animals were infected with a sheep-derived strain (SDS). Changes in the liveweights and some blood constitutents between the two infected groups were compared. The pathogenic effects of the GDS larvae were generally more serious when compared with those of the SDS larvae.     

Persistence of the efficacy of copper oxide wire particles against Haemonchus contortus in sheep

The aim was to determine the persistent efficacy of copper oxide wire particles (COWP) against Haemonchus contortus in sheep, using the harmonization guidelines protocol. Thirty-six male lambs (2 months old) reared free of gastrointestinal nematodes were used (average body weight of 10.8±3.8kg). Before and for the duration of the study, lambs were kept in raised cages with slatted floors and were offered ad libitum a complete mixed diet. Animals were divided into six groups (n=6): one non-treated control group (G0) and five groups treated with one COWP capsule (1.7g of copper oxide; Copinox(?)). Animals in each group were treated on pre-defined dates before the artificial infection was applied: days -35 (G1), -28 (G2), -21 (G3), -14 (G4) and -7 (G5). On day 0 animals were infected with 3700 H. contortus infective larvae per animal. Animals were humanely slaughtered between days 22 and 23 post-infection. The abomasums were individually washed to obtain the contents. These organs were subjected to separate artificial digestions. Adult parasites were counted from the abomasum contents and the larvae from the digested material. Worm burden geometric means were calculated for each group. A significant worm burden reduction in either of the treated groups (G1, G2, G3, G4, and G5) compared to the control (G0) was considered as persistence of the anthelmintic effect. Copper levels were determined from individual liver samples of each animal. The geometric mean worm burden of the control group (G0) was 1959. Compared to the control, worm burdens geometric means were significantly reduced in groups G1 (1108), G4 (528) and G5 (1063) (P<0.03). Efficacies in G1, G4 and G5 were 43.4%, 73.0% and 45.7% respectively. No significant reduction was found for G2 (1342) and G3 (1430). A larger quantity of Cu was found in the livers of treated animals compared to the control group (P<0.05) except for G3 (P=0.06). A negative association between Cu liver content and worm burdens was found (r=-0.42, P<0.05). Live weight gain was similar in all groups and no clinical or post-mortem manifestations of Cu toxicity were recorded in treated animals.     

Genetic variation of Haemonchus contortus (Trichostrongylidae) in sheep and goats from Malaysia and Yemen

The large stomach worm, Haemonchus contortus, commonly known as "the barber's pole worm", is a blood-sucking nematode found in the abomasa of sheep and goats. This work is the first documentation on the ND4 sequences of H. contortus from sheep and goats in Malaysia and Yemen and the results provide a preliminary insight on the genetic differences of H. contortus found in the two countries. In general, this study showed a high degree of diversity and low population structure of this species within the same country in comparison with higher genetic structuring at a wider geographical scale. The results also showed that the majority of genetic variance was within H. contortus populations. The Malaysian sheep and goat populations investigated appeared to share the same isolate of H. contortus while different isolates may be found in Yemen which must be taken into account in the design of an effective control strategy. Analysis of the internal transcribed spacer-2 (ITS-2) confirmed that all samples investigated in this study belonged to H. contortus. However presence of other Haemonchus species parasitizing these two hosts can only be confirmed by further detailed studies.     

捻转血矛线虫谷胱甘肽过氧化物酶体外对山羊外周血单核细胞功能的影响

本文探讨了捻转血矛线虫的谷胱甘肽过氧化物酶(HC29)体外对山羊外周血单核细胞(PBMCs)功能的影响。颈静脉采取无捻转血矛线虫感染的山羊血液,分离单核细胞后加入终浓度为5μg/mL的HC29重组蛋白进行体外培养,通过免疫荧光抗体技术观察重组蛋白与单核细胞的结合情况;HC29刺激山羊外周血单核细胞,用qPCR技术测定各试验组单核细胞中IL-2、IL-4、IL-10、IL-17、IFN-γ和TGF-βmRNA的转录水平;重组蛋白刺激单核细胞,采用CCK-8法测定细胞的增值情况;将重组蛋白与巨噬细胞共培养,测定巨噬细胞吞噬和分泌NO的情况。结果表明,重组HC29能够与外周血单核细胞结合并刺激细胞因子IL-10和IFN-γ的表达水平显著提高(P<0.01),能够引起单核细胞增值(P<0.01),巨噬细胞吞噬作用增强(P<0.01)和NO分泌增加(P<0.01)。结果表明HC29是捻转血矛线虫的一个重要抗原,主要诱导Th1类免疫反应。