Humoral immune responses of Hereford cattle vaccinated with midgut antigens of the cattle tick, Boophilus microplus

Vaccination of cattle with midgut membrane (GM) antigen derived from the cattle tick, Boophilus microplus, infected with the adjuvant Quil A, resulted in significant increases in total immunoglobulins, mainly in the IgG1 and IgG2 fractions of the serum. Analysis of the anti-GM antibody levels of vaccinated cattle showed that the levels of IgG, IgG1 and complement-fixing antibodies were significantly correlated to protection against infestation with cattle ticks. Anti-GM antibodies of the IgG2 and IgM isotype were not correlated to protection against infestation with cattle ticks. Anti-GM antibodies fixed complement (C') in the presence of GM, larval membrane antigen and live, midgut cells, but not in the presence of live, larval cells. Anti-GM antibodies were able to fix C' equally well in the presence of GM antigen and live, midgut cells. None of the antigens tested activated the alternate pathway of complement under the conditions tested. Levels of anti-GM IgG1 antibodies were used to develop a regression model for predicting levels of protection against infestation with cattle ticks in vaccinated cattle.     

Correlation between the infection rate of the vector tick, Haemaphysalis longicornis and the parasitaemia of cattle infected with Theileria sergenti

The infection rate of Theileria sergenti in salivary glands of nymphal Haemaphysalis longicornis which had dropped from cattle showing different piroplasm parasitaemias was examined by the methyl green-pyronin staining method. The results suggested that there was some correlation between parasitaemia and the subsequent infection rate in salivary glands of ticks. It seemed possible that the parasitaemia in cattle, especially the number of parasitized erythrocytes in blood imbibed by ticks, might be an important influence on the number of infected acini of ticks.     

Immune responses of cattle to biochemically modified antigens from the midgut of the cattle tick, Boophilus microplus

Treatment of membrane antigens of the midgut (GM) of the cattle tick, Boophilus microplus with sodium metaperiodate (periodate), pronase and lipase significantly inhibited the reactivity of the GM with antibodies in the sera of 57 cattle vaccinated with GM. Treatment of GM with periodate only removed the correlation between antibody reactivity of sera and protection against infestation with ticks. A monoclonal antibody (MoAb QU13), which recognises protective antigens solubilized from GM (Lee + Opdebeeck 1991), did not react with GM treated with periodate. Cattle vaccinated with GM extracts were significantly protected against infestation with cattle ticks (P less than 0.05), whereas cattle vaccinated with either GM extracts treated with periodate or with antigens precipitated from GM extracts with MoAb QU13 and also treated with periodate, were not protected against infestation. These studies provide preliminary evidence that protective antigens in the tick midgut membrane either are carbohydrate or are dependent on carbohydrate for their specificity.     

Larval membrane antigens protect Hereford cattle against infestation with Boophilus microplus

Hereford cattle (Bos taurus) were immunized with antigens solubilized with Triton X-100 from larval membranes of the cattle tick (Boophilus microplus). Based on tick egg production compared to control cattle, vaccinated cattle were protected (78%) against challenge with 2 x 20,000 tick larvae. The soluble Triton X-100 extract of tick larval membranes was further purified by immunoaffinity chromatography, using immunoglobulin ligands (IgG1 and IgG2) from three immune steers, previously vaccinated with membrane antigens from the midgut of partly engorged adult female ticks. Cattle vaccinated with these purified antigens were protected in two separate experiments (80 and 89% respectively), against challenge with 2 x 20,000 larval ticks compared to control cattle. Whole larval membranes used as vaccines in cattle reduced the amount of eggs produced from ticks by 47% compared to control cattle, but this difference was not significant.     

Hereford cattle immunized and protected against Boophilus microplus with soluble and membrane-associated antigens from the midgut of ticks

Hereford cattle were immunized with membranes and soluble components extracted from the midgut of Boophilus microplus. Membrane vaccines protected cattle (91%) against challenge with 3 x 20,000 larval ticks administered at intervals of 7 days. Vaccines made from soluble antigens did not protect cattle. Antibody levels measured by enzyme-linked immunosorbent assay (ELISA) related to the levels of protection induced by vaccination.     

Immunogenetic influences on tick resistance in African cattle with particular reference to trypanotolerant N'Dama (Bos taurus) and trypanosusceptible Gobra zebu (Bos indicus) cattle

In sub-Saharan Africa, tick infestation and tick-borne infections together with tsetse-transmitted trypanosomosis arguably constitute the main parasitological disease complex constraining livestock production. Resistance to tick attack and tick-borne micro-organisms (TBMs) varies among different breeds of cattle. The magnitude of losses due to these parasites is related to an extent to the degree of breed resistance. Generally, zebu (Bos indicus) cattle possess a higher resistance to ticks and TBMs than European (Bos taurus) cattle. The host's immune system would appear to be the single most important factor that regulates this resistance. This paper reports on the main effector immune mechanisms governing resistance against ticks and TBMs. The cellular immune response appears more effective and stable than humoral immunity in modulating resistance to ticks and TBMs. Similarities between the immune mechanisms employed by trypanotolerant N'Dama (B. taurus) cattle, when infected with trypanosomes, and those elicited by tick bites and TBMs seem to exist, particularly at the skin level in the early phases of parasitic invasion. Moreover, there is evidence that in the N'Dama breed, resistance against ticks per se also has a genetic basis. Therefore, the N'Dama appears to be a unique breed in that it exhibits resistance to several parasitic diseases and/or infections, including helminths, when compared to other cattle breeds in West Africa. It is concluded that the multi-parasite resistant traits of the N'Dama breed should be exploited in those areas where trypanosomosis, ticks and tick-borne diseases constrain animal production. This should be of benefit for low-input farming systems where the use of chemicals for prophylaxis and therapy is limited by their relatively high cost. Additionally, the potential contribution of multiple disease resistant N'Dama cattle should be considered in crossbreeding programmes with exotic dairy breeds for increasing milk production in West Africa.     

广西地区家畜牛、羊体表硬蜱情况调查及种类鉴定

目的 掌握广西地区牛羊场硬蜱种类及其分子特征,了解该地区蜱类的分类地位,为养殖户防控硬蜱及蜱媒传染病提供参考依据。方法 本实验于2019年1月至2021年11月,采用动物体表法采集寄生的蜱类;提取蜱基因组,PCR扩增3种硬蜱的线粒体16S rDNA和COI基因片段,并进行同源性分析。基于邻接法,用MEGA6.0软件分别构建系统发生树,进行遗传进化分析。结果 牛羊体表上共采集蜱2 030只,隶属1科2属3种,其中微小扇头蜱1 968只,长角血蜱49只,具角血蜱13只。PCR扩增微小扇头蜱、长角血蜱和具角血蜱16S rDNA和COI基因片段长度分别是460 bp和710 bp左右,分别与GenBank中已登录的相应种类相似较高且在一个进化分支上。结论 广西地区牛羊场优势蜱种是微小扇头蜱且存在长角血蜱和具角血蜱。三蜱种16S rDNA和COI序列存在多样性和地域差异性。     

Immunological responses and the role of the paralyzing toxin in rabbits infested with Rhipicephalus evertsi evertsi

Twenty-nine rabbits were exposed to 4 infestations with 20, 50, and 100 adult red-legged ticks (Rhipicephalus evertsi evertsi). Upon challenge, 16 rabbits (55%) contracted paralysis, and 14 of these (88%) died. Most deaths occurred in rabbits challenged with 100 ticks. Immunity to paralysis and subsequent resistance to tick feeding developed in the hosts that survived the initial challenge. Antibody titers to a salivary gland antigen increased progressively as determined by the following tests: passive hemagglutination, agar gel double immunodiffusion, and enzyme-linked immunosorbent assay. Rabbits challenged with 20 ticks displayed declining titers with advanced frequency of challenge and few of the rabbits in this group developed paralysis. There was no decline in antibody titers in groups of rabbits exposed to 50- and 100-tick loads. It is suggested that a certain quantum of antigen injected by the ticks is necessary for induction of immunity to paralysis.     

A survey of ticks on farm animals in Libya.

Thirteen species of ixodid ticks and two species of argasid ticks were collected during a three-year survey of 58 farms in Libya. These included Boophilus annulatus, B. microplus, B. decoloratus, seven species of Hyalomma, Rhipicephalus sanguineus, Rh. evertsi, Rh. bursa, Argas persicus and Ornithodoros foleyi. This is the first recording of B. microplus, B. decoloratus and Rh. bursa in Libya. Of 20,391 animals examined by random sampling, 2020 (9.6%) had ticks; particularly common were Hy. dromedarii on camels, Hy. impeltatum on sheep and Hy. excavatum on cattle. The tick found most frequently overall was Hy. dromedarii.     

Comparative efficacy of rHaa86 and rBm86 against Hyalomma anatolicum anatolicum and Rhipicephalus (Boophilus) microplus

Hyalomma anatolicum anatolicum and Rhipicephalus (Boophilus) microplus are the most economically important tick species in India and other tropical and subtropical regions of the world and transmit pathogens causing animal and human diseases. We demonstrated that vaccination of animal by rHaa86 could be used for the control of both H. a. anatolicum and R. (B.) microplus infestations. By comparing the efficacy of rHaa86 and rBm86, it was observed that vaccine based on rHaa86 will be more effective in controlling homologous challenge infestations (68·7% against larvae and 45·8% against adults). The results of this trial demonstrated that species-specific antigens are the better choice for vaccine development and could serve as an effective tool for the integrated control of H. a. anatolicum.     

Assessment of fungal isolates for development of a myco-acaricide for cattle tick control

Entomopathogenic fungal isolates of Arachnid origin were assessed for their ability to produce mortality and inhibit egg hatching in Boophilus microplus with the aim of selecting an isolate for development into a myco-acaricide for control of cattle ticks. The ability of the most promising isolate to target developmental stages of more than one tick species and the optimum concentration of fungal inoculum to be used for future studies were determined. Metarhizium anisopliae was the most pathogenic of the three fungal species tested on B. microplus, producing shorter average survival times (ASTs) for engorged adults (AST = 5.2 +/- 0.1 days) and larvae (AST = 9.3 +/- 0.4 days), and a longer average hatching times (AHT; AHT = 19.8 +/- 0.5 days) in comparison to Simplicillium lamellicola and Paecilomyces farinosus. In comparative studies on two tick species with similar life cycles, M. anisopliae produced a shorter AST in engorged adult B. microplus (AST = 8.8 +/- 0.3 days) than Rhipicephalus sanguineus (AST = 10.3 +/- 0.3 days). M. anisopliae was pathogenic to larvae of B. microplus (AST = 7.7 +/- 0.4 days), however, had no effect on larvae of R. sanguineus (AST = 14.6 +/- 0.3 days) as the AST of this treatment was similar to its untreated control (AST = 14.1 +/- 0.4 days). M. anisopliae lengthened the AHTs in both B. microplus (AHT = 16.4 +/- 0.3 days) and R. sanguineus (AHT = 16.7 +/- 0.3 days) in comparison to the controls. The ASTs of engorged adult B. microplus treated with M. anisopliae shortened as the concentration was increased from 1 x 10(7) to 5 x 10(8) conidia/ mL. A further increase in concentration, 1 x 10(9) conidia/mL (AST = 10.2 +/- 0.4 days) did not shorten or lengthen the AST in comparison to 5 x 10(8) conidia/mL (AST = 9.4 +/- 0.3 days).     

Adult Ixodes dammini on rabbits: development of acute inflammation in the skin and immune responses to salivary gland, midgut, and spirochetal components

Rabbits exposed to female Ixodes dammini (both uninfected and infected with Borrelia burgdorferi) or injected with B. burgdorferi showed an acute inflammatory response in the skin. Granulocytes and monocyte-histiocytes were the predominant infiltrating cells. Spirochetes were detected in the tick feeding cavities in the deep dermis. The inflammatory process was accompanied by polyclonal antibody responses to tick salivary gland components. Western blots showed that immune rabbit serum reacted with proteins of molecular masses of 8, 24, and 36-41 kilodaltons in both unengorged and engorged tick salivary glands. Additional reacting bands in the immunoblot of the engorged salivary gland indicated that new antigenic components of the salivary gland are synthesized during engorgement. Rabbits did not produce antibodies to tick midgut components. Murine monoclonal antibody 11G1 detected outer surface protein A of B. burgdorferi in immunoblots of midguts from unengorged ticks, faintly in engorged salivary gland, and seldomly in unengorged salivary gland, findings suggesting that the spirochete is transmitted to the host via tick saliva during the later stages of feeding.     

云南临沧地区蜱种分子生物学鉴定及埃立克体基因序列分析

目的 了解云南临沧地区蜱虫种类及其自然感染埃立克体的情况。方法 采集耕牛体表寄生的蜱虫,经形态学鉴定和分组后,从蜱虫中提取DNA,应用PCR扩增蜱虫COI基因以及埃立克体16S rRNA和groEL基因,并测序分析。结果 共采集到蜱虫42只,其中微小牛蜱38只占90.48%、血蜱4只占9.52%。将每种蜱的2只为1组,在21组样本中,检出COI片断21份(检出率100%);在相同的4组样本中均检出埃立克体16S rRNA片断和groEL片断4份(检出率19.05%)。基因序列分析,检出的COI片断序列中有2份与澳大利亚pava血蜱(JX573136)的同源性最高(89.3%),其余19份序列与马来西亚微小牛蜱B3的同源性最高(99.5%);检出的4份16S rRNA片断序列完全一致,与美国查菲埃立克体Arkansas和埃文埃立克体Aa2FT349及中国北京查菲埃立克体BY-YQ-HME-O18株的同源性均为100%;检出的4份groEL片断序列也完全一致,与日本埃立克体Yonaguni206 株的同源性最高均为93.9%。结论 经分子检测证实云南临沧地区耕牛体表微小牛蜱中存在一种类似日本Yonaguni206株的埃立克体感染,耕牛体表还可能寄生一种新的血蜱。     

Theileria mutans the infectivity for cattle of parasites derived from prefed Amblyomma variegatum nymphs.

A steer was infected with Theileria mutans (Aitong) by application of infected Amblyomma variegatum nymphs. Uninfected A. variegatum larvae were applied to this steer when piroplasms developed. The resultant (and infected) A. variegatum nymphs were applied to the ears of rabbits. Equal numbers of ticks were removed daily from rabbits and ground in Eagles Minimal Essential Medium. The infectivity of the supernatant fluid was tested in cattle. The produced supernatant fluids were infective between days 3 to 7 but noninfective on days 0, 1 and 2. The feeding of A. variegatum on cattle and rabbits was compared and the increase of haemoglobin in the supernatant was correlated with the weight increase of the nymphs. It is hoped that infective T. mutans (Aitong) stabilate derived from ticks can be produced using this information.     

Isolation and characterization of salivary antigens from Hyalomma anatolicum anatolicum

This study demonstrates the involvement of a large number of salivary proteins in the acquisition of resistance to Hyalomma anatolicum anatolicum. Using immunoblotting, sera from hypersensitized rabbits were shown to react with nine proteins in the saliva and 17 in salivary gland extracts (SGE) from 96 h fed female ticks. The salivary antigens had molecular weights in the range of 14 400 to 130 000. All the antigens identified in the saliva and 12 of the SGE antigens were glycoprotein in nature and a majority of them appeared to be common to different stages of feeding. In addition antigen I (molecular weight 130 000) showed acid phosphatase and antigen III (molecular weight 96 000) showed both non-specific esterase and aminopeptidase activity. Three high molecular weight proteins isolated from saliva (antigen I, antigen II--molecular weight 103 000 and antigen III), gave immediate hypersensitivity reactions in intradermal inoculation into rabbits which had previously been exposed to ticks. Antigens II and III also elicited a strong delayed hypersensitivity reaction. These results may help to explain the nature of the immune mechanisms which effect resistance against H. a. anatolicum.     

Tick-borne rickettsioses in Pune district, Maharashtra, India

An extensive study on tick-borne rickettsioses in the Pune district of Maharashtra revealed that Indian tick typhus exists as a zoonosis, which only occasionally causes disease in man. By sero-conversion in guinea pigs, presumptive isolates of Rickettsia conori and Coxiella burnetii were recovered from 4 of the 11 species of ticks examined. Boophilus microplus and Rhipicephalus haemaphysalis were found to be harbouring R. conori whereas C. burnetii was isolated from Haemaphysalis intermedia and Hyalomma hussaini in addition to the above mentioned 2 tick species. Complement fixation tests carried out on sera from various species of rodents and gerbils revealed the presence of antibodies against the R. conori antigen in the sera of Rattus blanfordi, R.r. rufescens and Suncus murinus. In the case of large mammals, similar antibodies were detected in the sera from dog, cow, horse and sheep. C. burnetii infection was found to exist in both the sylvan and domestic cycle, as evidenced from the involvement of ticks, large and small mammals and man in its natural history.     

Ixodid ticks on domestic animals in Samut Prakan Province,Thailand

The prevalence of tick-harboring domestic animals, tick density, and the species of ticks were studied throughout the year 2000, in Muang Samut Prakan, Bang Phli and Phra Pradaeng districts of Samut Prakan Province. The animals examined were Canis lupus familiaris (450), Bos indicus (cross-bred) (189), Bos taurus (30), Bubalus bubalis (171) and Sus scrofa domestica (450). The total number of collected ticks was 1,491. The pigs did not harbor ticks. The stages of ticks collected were larvae, nymphs and adults. The prevalence rates of tick-harboring were 46% (Canis lupus familiaris), 42.86% (Bos indicus, cross-bred), 33.33% (Bos taurus) and 9.35% (Bubalus bubalis). The tick densities were 2.22 (Bos indicus, cross-bred), 2.16 (Canis lupus familiaris), 1.16 (Bos taurus) and 0.36 (Bubalus bubalis). Only 2 species of ixodid ticks, Boophilus microplus and Rhipicephalus sanguineus, were found. R. sanguineus was the dominant species of tick. The percentage of R. sanguineus was 65.2% and B. microplus was 34.8%. In Muang district, R. sanguineus was the dominant species in C. lupus familiaris; in Bang Phli district, B. microplus was the dominant species in Bos indicus (cross-bred). The density of B. microplus was high in the summer season; the density of R. sanguineus was high in the winter season. The number of ticks depended on the geographic location, animal host and season.     

Immunoglobulin G binding proteins in male Rhipicephalus appendiculatus ticks

Three abundant immunoglobulin G binding proteins (IGBPs) of29kDa (IGBP-MA), 25kDa (IGBP-MB), and 21 kDa (IGBP-MC) were isolated from partly fed male Rhipicephalus appendiculatus ticks using an agar-ose-IgG column. After separating the proteins by SDS-PA GE, each individual IGBP band was cutou tfrom the gel and used to raise antiserum in guinea pigs. Using immuno-blotting, each of the three IGBPs was shown to be antigenically distinct, and specific for feeding male R. appendiculatus ticks. They were detected in the salivary glands of male ticks that had been feeding in either the presence of absence of female ticks, and appeared to be glycoproteins. The smallest protein, IGBP-MC, bound to guinea pig IgG (on which species the ticks were fed) and also bound to human and bovine IgG, whereas IGBP-MA and possibly IGBP-MB only bound to guinea pig IgG. The function of IGBPs in tick salivary glands and their significance in male ticks are discussed.     

Salivary antigens of the cat flea, Ctenocephalides felis felis

The cat flea, Ctenocephalides felis felis, is the major cause of flea bite hypersensitivity (FBH) in dogs and cats, yet little progress has been reported on identifying the antigens responsible. We obtained flea salivary antigens by washing secretions from containers probed by the mouthparts of fleas, and by extracting whole flea salivary glands. Mice were exposed to feeding fleas to generate antibodies to salivary antigens injected in vivo . The sera were tested for antibodies against the salivary antigens described and against a whole flea extract; in indirect ELISA, antibodies to salivary secretions were detected in 60% of the sera from mice exposed to feeding fleas. These sera identified four protein bands at apparent MW 56, 54, 42 and 40 K which corresponded to prominent protein bands in whole salivary gland extracts identified by protein staining after SDS-PAGE. Fixed sections of whole fleas exposed to the antisera showed that only structures within the salivary glands were identified. The salivary secretions and gland extracts are now being used to study immune responses of dogs suffering from FBH.