A Hypoderma diana (Diptera: Hypodermatidae) infection in a horse

An infection with second-stage larvae of the warble-fly H. diana in a horse is described. The second-stage larvae were incapable of developing into the third stage, because horses are unsuitable hosts and because the infected horse was treated with an insecticide. Since the horse was used for dragging trees in the forests, the infection was likely contracted via contact with H. diana, a normal parasite of roe deer in the Netherlands.     

First reported case of Hypoderma diana Brauer, 1985 (Diptera: Oestridae)-associated myiasis in a horse in Germany

A massive infestation with warble fly larvae was observed in a mare in Lower Saxony, Germany. From a herd of 20 horses, only a single mare accidently escaped the routine treatment with macrocyclic lactone in autumn and showed the myiasis infestation next year in January. Eighteen subcutaneous nodules were found on the back (n = 2), ribs (n = 3), flank (n = 5) and croup (n = 8). Deer, the specific hosts of the warble fly Hypoderma diana, was abundant on this farm and some other risk factors for the hypodermiosis are discussed herein. This is the first report of myiasis caused by H. diana in horses in Germany. This massive warble fly infestation in a horse in Germany reported here is important to alert equine health professionals concerning the prevention of this disease in the routine parasite control programme.     

Efficacy of injectable and pour-on microdose ivermectin in the treatment of goat warble fly infestation by Przhevalskiana silenus (Diptera, Oestridae)

The prophylactic efficacy of microdoses of injectable and pour-on ivermectin formulations against larval stages of Przhevalskiana silenus was assessed in naturally infected goats in the region of Calabria (southern Italy).Sixty-eight goats from two goat farms were divided into five groups: one group remained untreated, while the other four groups were treated with microdoses of ivermectin (5 and 10 microg/kg injectable formulation and 10 and 20 microg/kg pour-on formulation).The microdoses of ivermectin were fully effective in the treatment of goat warble fly infestation (GWFI) as no larvae emerged from the warbles in the treated groups, while all the larvae emerged in the control groups. Irrespective of the type of formulation used, the difference between the treated groups and the control group was statistically significant (P< 0.001). By contrast, no statistical differences were found between the goats treated with the injectable formulation and those receiving the pour-on applications, and between the two doses of the injectable and pour-on formulations used. Given the plasma concentrations it attains at its lowest dose (0.052 - 0.042 ng/ml for the injectable formulation and 0.030 ng/ml for the pour-on) the injectable formulation seems to offer the most reliable route for the administration of ivermectin microdoses and it is acceptable for milk consumption. The introduction of ivermectin in the early eighties and the use of microdoses in some cases have made it possible to control cattle hypodermosis in large areas of Europe. As with cattle hypodermosis, the administration of ivermectin microdoses in goats is particularly interesting because of the low costs involved and the low levels of residues found in goat milk; it may thus constitute the basis for GWFI control campaigns in areas where the disease is prevalent.     

Cross-transmission studies with Hypoderma lineatum de Vill. (Diptera: Oestridae): attempted infestation of goats (Capra hircus)

The potential for cross-transmission of Hypoderma lineatum from cattle to domestic goats (Capra hircus) was examined using artificial infestation techniques. Two routes of infestation, subcutaneous injection and dermal penetration, were used to expose goats to newly hatched first instars. Presence of antibodies and appearance of circulating antigen (hypodermin C) were evaluated at selected intervals for up to 40 weeks post-infestation. In addition, immunoblots against H. lineatum first-instar proteins were conducted using sera taken at 10 weeks post-infestation. Goats were palpated for the presence of developing larvae at sub-dermal sites beginning at week 30 pi. No developing larvae were palpated at any time, regardless of the route of infestation nor was circulating antigen detected in any infested goats. Antibodies were present at weeks 6 and 10 and week 27 pi in both infested groups. Immunoblots indicated all infested goats produced antibodies to first instar H. lineatum antigens. H. lineatum appears to be incapable of completing development in domestic goats although the transient appearance of ELISA detectable antibodies and the presence of bands on immunoblots suggests that at least some larvae survive long-enough to engender a humoural response. The host specificity of H. lineatum is discussed in light of the general concepts of host-parasite relationships of oestrids.     

Serodiagnosis of goat warble fly infestation by Przhevalskiana silenus with a commercial ELISA kit.

Cross-reactivity between Hypoderma lineatum antigen and anti-Przhevalskiana silenus antibodies has been demonstrated by an ELISA technique. To evaluate the applicability of a commercial ELISA kit for the immunodiagnosis of goat warble fly infestation, different dilutions of serum and conjugate were tested, the development of antibody to P silenus in naturally infested goats was studied, and the results were compared with an ELISA technique using an antigen extracted from the first instar larvae of H lineatum. The best results were obtained with a serum dilution of 1:50; with both techniques the highest antibody concentration were recorded in October, November and December. In view of the confirmed cross-reactivity between H lineatum antigen and anti-P silenus antibodies, and the simplicity and rapidity of the assay, the commercial ELISA kit can be considered as a useful tool for the diagnosis of goat warble fly infestation.     

Impact of previous infestation on dynamics of circulating hypodermin C in cattle artificially infested with Hypoderma lineatum (Diptera: Oestridae)

Four groups of cattle were artificially infested with 50 first instar Hypoderma lineatum after either a primary natural infestation or in the absence of a primary infestation. In two groups the primary infestation had been terminated by the application of either an organophosphate insecticide or a macrocyclic lactone parasiticide. Circulating hypodermin C and specific antibodies were measured for 40 weeks after the artificial infestation. Stage specific mortality of the larvae was also monitored. Previously uninfested cattle exhibited typical antibody and antigen profiles during the infestations. Antibodies were first detected on Week 7 p.i., they rose to maximum values between Weeks 24 and 25, then declined as larvae reached the back. A second peak occurred at Week 32 and antibody remained more less constant thereafter. Previously infested groups exhibited a dramatic anamnestic response by Week 3 p.i. with antibody levels peaking at Week 8. A second peak was noted between Weeks 24 and 26 p.i. after which antibody levels declined and then remained relatively stable. The dynamics of circulating hypodermin C in the previously infested cattle resembled those in the previously uninfested cattle. Mortality of first instars did not differ among the four groups. Similarly mortality of second and third instars, in the warble, did not differ although there was a tendency for higher mortality in the previously infested, untreated group. These results reinforce previous work demonstrating the development of a significant immune response during the primary infestation that is reflected in the rapid and substantial production of antibodies upon re-infestation. It is significant that a challenge model using subcutaneous injection of newly hatched first instars avoids host immune responses in the skin that result in substantial larval mortality. The current data also support the concept that migrating first instars induce significant reduction in host immune response. There is a peak of antibody production as antigen levels peak during first instar migration to the back. As larvae molt to the second instar and antigen production ceases there is a persistence of antibody which suggests release of the immune response from the suppression induced by the first instar secretory antigens.     

Molecular differentiation of Hypoderma bovis and Hypoderma lineatum (Diptera,Oestridae) by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)

The most variable region of the cytochrome oxidase I (COI) gene of Hypoderma bovis(1) and Hypoderma lineatum(2) (Diptera, Oestridae) was amplified by PCR and the amplicons were sequenced and analysed. PCR products were digested with three restriction enzymes, namely BfaI, HinfI and TaqI, providing informative profiles. H. bovis and H. lineatum sequences revealed an inter-specific variation rate of 8.5%, and an intra-specific variation rate of 0.87 and 0.29%, respectively. The results showed that the COI gene region examined was useful for the differentiation of H. bovis and H. lineatum and that a PCR-RFLP assay is a practical tool for their identification, offering additional diagnostic and epidemiological instruments for the study of cattle grub infestation.     

Persistent activity of topical ivermectin against artificial infestations with Hypoderma lineatum (Diptera: Oestridae)

In controlled experiments utilizing artificial infestations, a topical formulation of ivermectin (IVOMEC Pour-On for Cattle) was 100% effective against migrating first instar Hypoderma lineatum for 3 weeks following treatment. Larvae were apparently killed early in the infestation as no significant level of specific antibodies was found in the treated calves. At 4 weeks post-treatment the efficacy remained high at 96%; mortality of larvae in the one calf in which warbles were observed and from which mature third instars were collected, was higher than that seen in the untreated calves, indicating some level of treatment induced effect. No specific antibodies were detected in calves that did not develop palpable warbles. Antibody kinetics in those calves from which viable larvae emerged were typical. The length of activity of this product against early stages of the cattle grub makes it practical to apply treatment up to 3 weeks before the end of fly activity.     

Degradation of bovine C3 by serine proteases from parasites Hypoderma lineatum (Diptera, Oestridae)

Purified enzymes of Hypoderma lineatum (Insecta, Oestridae), were assayed for their proteolytic activity on bovine C3 in normal cattle sera. The products of cleavage by these serine proteases (hypodermins A, B, and C), were analysed by electrophoresis in SDS polyacrylamide gels followed by immunoblotting. The enzymatic attack was initially directed at the alpha polypeptide chain by hypodermin A at a concentration of 1 μg/ml of serum and by hypodermin B at 5 μg/ml. The generated peptides differed in their molecular size from those produced during natural degradation of C3 in a control serum by physiologically relevant enzymes. Hypodermin A, at a concentration of 1 μg/ml, also caused a cleavage of the β chain. At 5 μg/ml, hypodermin A induced total degradation of the C3 molecule. Hypodermin B (5 μg/ml) starts splitting C3 near cleavage sites of factor I. Bovine C3 appears to be highly sensitive to hypodermins A and B in normal sera. Apparent molecular sizes and alignment of the bovine C3 cleavage products are presented schematically. Hypodermin C, a collagenolytic enzyme, had no effect on C3 in normal sera. The biological consequences for the immunopathological reactions associated with hypodermosis are discussed.     

Tick infestation of sika deer (Cervus nippon) in the western part of Yamaguchi Prefecture,Japan

Ticks were collected from 94 sika deer (Cervus nippon) hunted in the western part of Yamaguchi Prefecture, Japan from August to November 1999, and March to July 2000. Haemaphysalis longicornis and H. yeni were the dominant species from April to August, while H. flava and H. megaspinosa were dominant in October, November and March. This is the first report of H. yeni in the mainland of Japan. Small numbers of H. kitaokai, Amblyomma testudinarium and Ixodes ovatus were also recorded.     

The moose throat bot fly Cephenemyia ulrichii larvae (Diptera: Oestridae) found developing in roe deer (Capreolus capreolus) for the first time

About fifty larvae of Cephenemyia ulrichii Brauer (Diptera: Oestridae), some of them nearly full-grown third instars, were found in the throat of a roe deer (Capreolus capreolus) in June 2007 near Helsinki in Finland. The parasite is considered to be host specific, occurring only in the moose (Alces alces), and this paper is apparently the first report of a successful infestation in an aberrant host.     

驯鹿的皮蝇蛆线粒体CO1基因序列分析

利用分子生物学技术对内蒙古驯鹿未定种皮蝇蛆线粒体CO1基因种属特异性序列进行了研究.DNA核苷酸测序结果证实:该种皮蝇线粒体CO1种属特异性基因UEA7到UEA10特殊目标编码区域片段长度约为689 bp;种系发生进化树和同源性分析显示其与鹿皮蝇法国株同源性非常接近,因此确定内蒙古地区感染驯鹿的皮蝇蛆为鹿皮蝇(Hypo...     

Chrysomya bezziana (Diptera: Calliphoridae) infestation in two Malaysian cats treated with oral lotilaner

The most common fly species associated with screwworm myiasis in Southeast Asia is Chrysomya bezziana (Ch. bezziana), the Old‐World screwworm. Treatment of screwworm myiasis in cats traditionally has comprised subcutaneous injection of ivermectin or oral administration of nitenpyram, combined with aggressive tissue debridement and larval removal under general anaesthesia. Two cats diagnosed with cutaneous myiasis caused by the larvae of Ch. bezziana were treated with lotilaner. In both cats, a single dose of lotilaner at 6–26 mg/kg, killed all larvae within 24 h, negating the need for general anaesthesia. Both cats were simultaneously infested with Lynxacarus radovskyi (L. radovskyi) which also was eradicated with lotilaner. No adverse reactions were observed and both cats recovered without complications.     

Effect of treatment on the dynamics of circulating hypodermin C in cattle naturally infested with Hypoderma lineatum (Diptera: Oestridae)

An antigen capture ELISA, using a murine monoclonal antibody recognising recombinant hypodermin C (rHyC), was used to evaluate the influence of early treatment with eprinomectin (Eprinex) or fenthion (Spotton) on the kinetics of circulating hypodermin C in calves naturally infested with Hypoderma lineatum. No viable larvae were collected from treated animals, whereas a variable number of warbles were found in control animals. Treatment provoked a decrease in circulating HyC levels that was significant 9 days post-treatment (p.t.). Circulating antigen levels in the treated cattle remained detectable for approximately 99 days p.t. In contrast, control animals had no detectable antigen at 64 days p.t., 42 days earlier than in the treated animals. These results suggest that larvae were either gradually killed, resulting in slow release of antigen or they were encapsulated, leading to the slow liberation of antigen. Kinetics of circulating HyC did not differ among the two insecticide treatments. Antibodies persisted, in all groups, throughout the 120-day study. These results suggest that the antigen capture ELISA will be useful as a technique for detecting successful treatment of cattle grub infestations and for the detection of new infestations in previously infested cattle.