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Ultrastructure of the integument of first-instar Hypoderma lineatum and H. bovis (Diptera: Oestridae).

The ultrastructure of the integument of newly hatched, 60-d-old (in vitro-cultured) and 6-7-mo-old (recovered from the host) first-instar cattle grubs, Hypoderma lineatum (Villers) and H. bovis (L.), was studied. Changes in the cuticle during development of the first instar included increased thickness of the procuticle (0.8-1.4 to 7.5-7.9 microns) with an accompanying increase in number of lamellae (4-7 to 35-40), increased thickness of epicuticle (0.11-0.24 to 0.92 microns), and development of new epicuticle layers. The epithelial cells in newly hatched larvae also changed with age when compared with those in culture and those recovered from hosts. These changes included the appearance of dense plaques on the apical cell membrane and an increase in the folding of the basal cell membranes, with an accompanying increase in the amount of extracellular space. The basement membrane, separating epithelial cells from the hemocoel, also increased in thickness during the first instar. These changes reflect developments required to accommodate increased size but, in the case of the epithelial cells, suggest altered function. Ruthenium red staining on the cuticular surface of larvae cultured in vitro was indicative of the presence of negatively charged molecules, which may influence the host recognition and response capabilities.

Animals↗

New procedures to enhance survival of third-instar Hypoderma lineatum (Villers) (Diptera: Oestridae) in artificial media.

New techniques for the maturation of late third instars of the common cattle grub, Hypoderma lineatum (Villers), in artificial media are described. Larvae were held in either 24-well culture plates with media plus penicillin, streptomycin sulfate, nystatin, and chloramphenicol or in small salve jars on Perlite and media plus the same antibiotics. Chloramphenicol was not always necessary in the jars. Survival to pupariation of young third instars increased from 5% without nystatin and chloramphenicol to 73% when the two antibiotics were present in the culture plates. The survival of mature third instats to pupariation increased from approximately 53 to 80% after addition of nystati and chloramphenicol to the culture plates. Survival to pupariation of more mature grubs was similar in the jar and culture plate techniques. The former was more convenient, but the young grubs did not survive well in the jars.

Abattoirs↗

Method for collecting Hypoderma spp. (Diptera: Oestridae) larvae from cattle.

A new method for collecting 3rd-instar Hypoderma lineatum (Villers) and Hypoderma bovis (L.) is described. A procedure for carrying out chronobiological and other studies involving the life cycle of these flies is included. The larvae are collected from cattle using a dressing made of a circle of cardboard attached to a piece of tulle similar in color to the coat of the animal. These small cloth cages were attached to the animal skin with fast-drying contact glue wherever the swellings produced by 2nd- or 3rd-instar Hypoderma spp. had been previously detected. Each animal was inspected daily to detect the appearance of new larvae. This procedure allows the collection of each emerged larva without contact with other larvae and avoids friction of the cloth cages or dressing with the larvae, thus reducing larval death and increasing viability of the parasite.

Animals↗

Eosinophilic pleural effusion in cutaneous myiasis.

We have reported the case of a 54-year-old man with recurrent painful migratory subcutaneous nodules associated with marked blood eosinophilia and an eosinophilic pleural effusion. The entire syndrome was subsequently determined to be due to cutaneous myiasis caused by the larvae of Hypoderma lineatum, the cattle botfly. Infestation by this or other dipterous fly larvae should be among the parasitic diseases considered in the differential diagnosis of any patient with similar symptoms.

Diagnosis, Differential↗

Scanning electron microscopy of the posterior spiracles of cattle grubs Hypoderma bovis and Hypoderma lineatum.

Posterior spiracles of newly hatched first instar larvae of Hypoderma bovis (L.) and H. lineatum (DeVill.) consist of two pairs of spiracular openings. Each pair is surrounded by a rima bearing three spines. Posterior spiracles of second instar larvae are composed of a pair of medial ecdysial scars bounded laterally by spiracular plates. H. bovis spiracular plates have twenty-nine to forty openings, each surrounded by a slightly raised rima. H. lineatum spiracular plates have eighteen to twenty-five openings. Spiracular openings lead to posterior felt chambers which are connected to a common anterior felt chamber filled with a meshlike network. In third instar H. bovis each medial ecdysial scar is surrounded by a strongly concave spiracular plate. Spiracular openings are surrounded by slightly raised rima. Most rimae bear a spine. Spiracular plates of H. lineatum are flat and rimae are without spines. Each spiracular opening leads to a posterior felt chamber, several of which are confluent with a larger anterior felt chamber. Anterior felt chambers open into the dorsal longitudinal tracheal trunk. Felt chambers in third instar larvae are also filled with a complex mesh.

Animals↗

Early detection of cattle grub (Hypoderma lineatum and H. bovis) (Diptera, Oestridae) using ELISA.

An enzyme-linked immunosorbent assay (ELISA) to detect cattle grub-infested animals in the autumn in Alberta has been developed. Antibody to Hypoderma lineatum de Vill. was detectable as early as 6 weeks post-infestation in artificially infested steers. Peak antibody concentrations preceded the peak in maximum 'apparent' grub numbers which occurred between 37 and 43 weeks after infestation. Natural infestations with H. lineatum and H. bovis L., ranging from one to ninety-two grubs per calf, were readily detected by November and the incidence of false positives in uninfested calves was only 5%. Low level (1-4 grubs) infestations of H. bovis only became detectable in February with peak antibody concentrations occurring at the end of April. Prevalence of cattle grub infestations in southern Alberta was shown to be 37% during this study.

Animals↗

Preliminary evaluation of four immunological tests for the early diagnosis of Hypoderma tarandi causing hypodermosis in reindeer.

294 serum samples from five Norwegian reindeer herds were examined for antibodies against Hypoderma tarandi L. The first and second larval instars of H. tarandi were tested as antigens in immunodiffusion tests, passive haemagglutination and an Enzyme-Linked Immunosorbent Assay (ELISA). The latter technique, using first instar antigens, produced the best results. A significant difference (P less than 0.1%) was observed between the antibody value of naive reindeer bred in France and those from infected Norwegian herds. No correlation was observed between the antibody titre and the number of warbles recovered at necropsy from the 294 Norwegian reindeer.

Animals↗

Hypodermosis in the red deer Cervus elaphus in Córdoba, Spain.

The prevalence of red deer hypodermosis and the life cycle of Hypodermia diana Brauer 1858 in three provinces of the south of Spain, Córdoba, Jaen and Ciudad Real, were studied by inspecting 254, thirty-eight and thirty-five deer from each province respectively. The prevalence of infestations was: Córdoba, 87.75%; Jaen, 92.10%, Ciudad Real, 91.42%. From this we deduced an overall prevalence of 88.67%, comprising 88.23% in male deer and 89.96% in female deer. The intensity of the parasitism in 213 of the 245 animals from Córdoba varied between one warble per animal to more than 100 in 28 of the inspected animals. The chronology of the life cycle during the hunting period (October to the end of February) was investigated.

Animals↗

Changes in the haemolytic activity of bovine serum complement by Hypoderma lineatum (insect oestridae) larval proteinases in naive and immune cattle.

Three serine proteinases (hypodermin A, B and C) of the first instar larvae of Hypoderma lineatum have been assayed for their ability to deplete seric complement of naive or immune cattle. In naive cattle complement consumption is initiated by hypodermin B through the sequence C1-C3 at a concentration of 5 micrograms/ml of serum, and by hypodermin A through the sequence of C3-C9 at a higher concentration of 150 micrograms. The third enzyme presenting a collagenolytic activity has no anti-complementary activity even on C1q. In immune cattle a 70% complement depletion through the classical pathway is observed with 15 micrograms of hypodermin B per ml of serum. The two enzymes A and C appear to play a minor role in the complement depletion via the classical pathway. The biological role of each of these enzymes in the parasite-host interrelationships is discussed. The participation of these enzymes in the immediate hypersensitive reactions following systemic treatment of cattle infested by this endoparasite is considered.

Animals↗

Ophthalmomyiasis caused by the reindeer warble fly larva.

Two boys with ophthalmomyiasis caused by the first instar larva of the reindeer warble fly Hypoderma tarandi are reported. Both were 9 years old and came from the coast of northern Norway. One had ophthalmomyiasis interna posterior and one eye had been removed because of progressive pain and blindness. Histological examination showed the remains of a fly larva. The second boy had ophthalmomyiasis externa with a tumour in the upper eyelid, and histological examination showed a warble with a well preserved larva. Identification of the parasite in the histological material was based on the finding of cuticular spines and parts of the cephalopharyngeal skeleton identical with those of the first instar larva of H tarandi.

Animals↗