Ecology of Thelazia spp. in cattle and their vectors in Italy

The genus Thelazia (Spirurida, Thelaziidae) includes a cosmopolitan group of eyeworm spirurids responsible for ocular infections in domestic and wild animals and transmitted by different species of muscids. Bovine thelaziosis is caused by Thelazia rhodesi Desmarest 1828, Thelazia gulosa Railliet & Henry 1910, and Thelazia skrjabini Erschow 1928, which occur in many countries; T. gulosa and T. skrjabini have been reported mainly in the New World, while T. rhodesi is particularly common in the Old World. In Italy, T. rhodesi was reported in southern regions a long time ago and, recently, T. gulosa and T. skrjabini have been identified in autochthonous cattle first in Apulia and then in Sardinia. Thirteen species of Musca are listed as intermediate hosts of eyeworms, but only Musca autumnalis and Musca larvipara have been demonstrated to act as vectors of Thelazia in the ex-URSS, North America, ex-Czechoslovakia and more recently in Sweden. In Italy, after the reports of T. gulosa and T. skrjabini in southern regions, the intermediate hosts of bovine eyeworms were initially only suspected as the predominant secretophagous Muscidae collected from the periocular region of cattle with thelaziosis were the face flies, M. autumnalis and M. larvipara, followed by Musca osiris, Musca tempestiva and Musca domestica. The well-known constraints in the identification of immature eyeworms to species by fly dissection and also the time-consuming techniques used constitute important obstacles to epidemiological field studies (i.e. vector identification and/or role, prevalence and pattern of infection in flies, etc.). Molecular studies have recently permitted to further investigations into this area. A PCR-RFLP analysis of the ribosomal ITS-1 sequence was developed to differentiate the 3 species of Thelazia (i.e. T. gulosa, T. rhodesi and T. skrjabini) found in Italy, then a molecular epidemiological survey has recently been carried out in field conditions throughout five seasons of fly activity and has identified the role of M. autumnalis, M. larvipara, M. osiris and M. domestica as vectors of T. gulosa and of M. autumnalis and M. larvipara of T. rhodesi. Moreover, M. osiris was described, for the first time, to act as a vector of T. gulosa and M. larvipara of T. gulosa and T. rhodesi. The mean prevalence in the fly population examined was found to be 2.86%. The molecular techniques have opened new perspectives for further research on the ecology and epidemiology not only of Thelazia in cattle but also of other autochthonous species of Thelazia which have been also recorded in Italy, such as Thelazia callipaeda, which is responsible for human and canid ocular infection and Thelazia lacrymalis, the horse eyeworm whose epidemiological molecular studies are in progress.     

Bovine thelaziasis in Iowa

The prevalence of developing Thelazia nematodes in face flies (Musca autumnalis) was studied for 7 yr at a beef farm in central Iowa. Juvenile nematodes were not found among flies in reproductive diapause in autumn, nor among nulliparous, overwintered flies in spring, but only among actively reproducing insects. Thus Thelazia probably do not overwinter in face flies. A mean prevalence of 2.0% infected was recorded among flies in 7 fly breeding seasons. No heterogeneity in Thelazia prevalence was detected within fly breeding seasons. The frequency distribution of Thelazia among face flies by year of occurrence was homogeneous, with a mean of 2.75 larval nematodes per infected fly. Thelazia gulosa and T. skrjabini were recovered in necropsy from the eyes of bovines from central Iowa. Prevalence among fat cattle 18-27 mo old was 15%; among cows 3-15 yr old, prevalence was only 3%. Thelazia skrjabini was found in 29 eyes and T. gulosa in 4 eyes. Infections were randomly distributed among the eyes of subject cattle. The mean worm burden was 2.3 T. skrjabini and 12 T. gulosa per infected eye. Thelaziasis is clearly enzootic in Iowa.     

Molecular characterization of the first internal transcribed spacer of ribosomal DNA of the most common species of eyeworms (Thelazioidea: Thelazia)

Spirurid nematodes of the genus Thelazia cause ocular infection in several mammals and are transmitted by dipteran flies. Of the 10 described species, T. gulosa, T. rhodesi, T. skrjabini (affecting cattle), T. lacrymalis (affecting horses), and T. callipaeda (affecting carnivores and humans) are the most common. The aim of this study was to characterize the first internal transcribed spacer (ITS1) ribosomal DNA sequences of these 5 species. The length of the ITS1 sequences ranged from 357 bp (T. lacrymalis) to 905 bp (T. callipaeda). Interspecific differences ranged from 35 to 77%. Polymorphic sites were detected in each species. Intraspecific variation varied from 0.3 to 2.5%. The characteristics of the ITS1 of Thelazia spp. show similarities to those of other Spirurida. The findings of this present study show that the ITS1 represent a powerful genetic marker for the molecular identification of eyeworms and a useful tool for developing molecular epidemiological techniques to study Thelazia spp. transmission patterns and prevalence in definitive and intermediate hosts.     

A study on the prevalence and intensity of occurrence of Thelazia skrjabini (Nematoda: Thelazioidea) in cattle in central Alberta, Canada

The distribution of Thelazia skrjabini in the eyes of cattle was examined from necropsies on 297 animals. Although the overall prevalence in beef cattle (21.5%) was similar to that in dairy cattle (25.7%), in general worms were more abundant in beef than in dairy animals. The worm prevalence was lowest in the months from March to June and highest in September among beef and dairy animals. Thelazia skrjabini was found in 76 (16.8%) eyes from beef, and 23 (16.4%) eyes from dairy cattle. Infections were not randomly distributed among the eyes of cattle. Significantly more dairy and beef cattle contained T. skrjabini in both eyes than expected based on the observed prevalences. Based on chi-square probabilities, significantly more cattle more than 2 yr of age and less than 10 mo of age were infected with T. skrjabini. Although all age groups were infected, more worms were observed in cattle more than 2 yr of age. More female worms than males were collected from cattle. The male-to-female ratio of 1:2 was consistent regardless of cattle age, season, or type.     

Prevalence and intensity of Thelazia spp. (Nematoda: Thelazioidea) in a Musca autumnalis (Diptera: Muscidae) population from Central Alberta

A face fly (Musca autumnalis) population near Wetaskiwin, central Alberta, Canada, was sampled 9 times from 26 July to 29 September 1988 for the early larval stages of Thelazia spp. Of 426 female flies examined, 159 (37%) contained Thelazia spp. (almost exclusively T. skrjabini), with an average worm burden of 4.2 larvae per infected fly. Prevalence ranged from 17 to 56% over 9 collections. This is the first report of Thelazia skrjabini in flies from western North America and the highest Thelazia prevalence in face flies yet reported in North America. The face fly population was also parasitized by Heterotylenchus autumnalis, with a prevalence of 5.5%.     

Musca osiris--a new intermediate host of Thelazia gulosa

Musca osiris Wd. was first recorded as an intermediate host of the nematode Th. gulosa. In Tadjikistan Musca autumnalis Deg. was first recorded as an intermediate host of Th. gulosa and Th. skrjabini. The flies are suggested to be used as indicators of thelaziasis infection by using a key suggested for the first time for the definition of species of larvae belonging to the genus Thelazia during differentiation of infected larvae from flies.     

Occurrence of Thelazia lacrymalis (Nematoda, Spirurida, Thelaziidae) in native horses in Italy

A survey on the prevalence of Thelazia spp. in the province of Bari (Apulia region, Italy) in slaughtered native horses was conducted from June 20, 1995 to April 3, 1996. Both eyes from 409 ten-month- to 4-year-old native animals were examined. Sixty horses (14.7%) were found parasitized by Thelazia lacrymalis. Three hundred-sixty one parasite specimens (220 females, 99 males and 42 larvae) were collected with a mean count burden of 6.0-5.1 (range 1 to 20) per head. T. lacrymalis specimens were found free in the conjunctiva and behind the nictitancte, in the excretory ducts of the Harderian gland and in the ducts of the lacrimal glands. Thelazia specimens were significantly more common in horses of 3-4 years. Gross examination of all infected animals showed a follicular conjunctivitis. This is the first report of T. lacrymalis horse infection in Italy.     

Occurrence of Thelazia lacrymalis (Nematoda, Spirurida, Thelaziidae) in native horses in Abruzzo region (central eastern Italy)

A survey on the prevalence of Thelazia spp. in Abruzzo region (Italy) in slaughtered native horses was conducted from August 29, 1997 to August 28, 1998. Both eyes from 128 eight-month to 11 year-old native animals were examined. 50 horses (39.06%) were found parasitized by Thelazia lacrymalis. 502 specimens (371 females, 88 males and 12 larvae) were collected. In the infected horses the numbers of T. lacrymalis ranged from 1 to 48, with a mean count burden of 3.92 per head (SD = 7.79). T. lacrymalis specimens were mainly in the excretory ducts of the Harderian gland, and also in the ducts of the lacrimal glands, free in the conjunctiva and behind the nictitancte. Gross examination showed a conjunctivitis, more frequently a follicular conjunctivitis, in the 58% of the infected horses.     

Fauna and distribution of nematodes from the suborders spirurata and filariata parasitizing cattle in Dagestan, from the perspective of vertical zoning

Cattle of Dagestan are infested with several nematode species from the suborders Spirurata and Filariata, and extensiveness of the invasion depends greatly on the altitude above sea level. Level of infestation with Thelazia rhodesi, Th. gulosa, and Th. skrjabini is 38% in plains, 20% in submontane, and 5% in mountain zone. The same tendency is observed for the species Gongylonema pulchrum--45, 22, and 10%, respectively. Infestation with Setaria labiato-papillosa is 27.3% on average. Infestation with Onchocerca gutturosa and O. lienalis is 11% in plains and 3% in mountain and submontane zones; infestation with Stephanofilaria assamensis and S. stilesi in these zones is 18 and 5%, respectively. In mountain localities situated higher than 1000 m a. s. l. only G. pulchrum is occurred.     

Thelazia lacrymalis in horses in Kentucky and observations on the face fly (Musca autumnalis) as a probable intermediate host.

Eyes from 114 (30.3%) of 376 dead horses, examined from 3 April 1975 to 3 April 1976, were naturally infected with adult Thelazia lacrymalis; 1 horse was also infected with 1 male Thelazia skrijabini. Adult T. lacrymalis from dead horses were successfully transferred mechanically to the eyes of 3 of 4 Shetland ponies raised helminth-free. Larvae from gravid female T. lacrymalis underwent development in experimentally infected, laboratory-raised face flies (Musca autumnalis) and third-stage larvae ranging from 1.82 to 2.94 mm in total length were recovered at 12 to 15 days postexposure. A total of 866 naturally occurring face flies were collected from the head region of horses. Twelve of the face flies harbored larval stages of Thelazia spp. One of the larvae resembled third-stage T. lacrymalis that were recovered from the experimentally infected, laboratory-raised face flies. Introduction of 3 third-stage larvae from 1 face fly onto the cornea of a pony raised helminth-free resulted in the recovery of 1 male T. skrjabini 242 days later. In addition to the eyeworm larvae, other parasites recovered from the face flies included Heterotylenchus autumnalis, hypopi of astigmatid mites and a first instar beetle (Coleoptera: Rhipiphoridae). Data from these investigations indicate the likelihood that face flies are an intermediate host for T. lacrymalis and probably other species of Thelazia in this part of the country.     

Thelazia rhodesi in a dairy farm in Romania and successful treatment using eprinomectin

Bovine thelaziosis can be caused by Thelazia rhodesi Desmarest 1828, Thelazia gulosa Railliet & Henry 1910 and Thelazia skrjabini Erschow 1928 which are known to be transmitted by species of Muscidae. Therapeutic measures in cattle include mechanical removal of the adult parasites, followed by irrigation of the conjunctival sac with different solutions such as levamisole (1%) or administration of ivermectin (0.2 mg/kg SC), doramectin (0.2 mg/kg SC or pour-on) both with high efficacy, however, the use of such macrocyclic lactones can be problematic in lactating cattle. Nematodes harboured in the eyes of some cattle from Romania were morphologically identified as Thelazia rhodesi and the PCR analysis confirmed the diagnosis with a 98.0% nucleotide similarity for ITS1 sequence, with other sequences available in GenBank. The present paper reports the presence of T. rhodesi in cattle from a dairy farm in Romania and the recovery after treatment with eprinomectin. To the best of our knowledge this is the first report on the efficacy of eprinomectin against Thelazia spp. and the first molecular confirmation of T. rhodesi in Romania.     

Development of the nematode eyeworm, Thelazia skrjabini (Nematoda: Thelazioidea), in experimentally infected face flies, Musca autumnalis (Diptera: Muscidae)

The development of Thelazia skrjabini Erschow, 1928, was studied in experimentally infected laboratory-reared Musca autumnalis De Geer. Thelazia skrjabini developed to the infective third stage in a minimum of 9 days in M. autumnalis maintained at 27 +/- 2 C. First-stage larvae were not observed postinoculation, but second-stage larvae were first observed 3 days postinoculation. Development was asynchronous. Second- and third-stage larvae occur in capsules, occasionally in the head but primarily in the abdomen attached to fat bodies. First-stage larvae have anteriorly 1 ventral and 2 dorsal hooks, directed posteriorly. Second-stage larvae have 4 submedian cephalic papillae and faint annular striations. Third-stage larvae have 6 labial papillae, 4 submedian cephalic papillae and pronounced annulations. Morphometric studies of each larval stage were performed with specimens in glycerine.     

Enzooticity of the dogs,the reservoir host of Thelazia callipaeda,in Korea

The reservoir hosts of Thelazia callipaeda were examined. The eyes of the 76 dogs raised at farm. 78 military dogs (shepherds), 96 cattle, and 105 pigs were investigated for the presence of eyeworm. Among them, six worms of T. callipaeda were collected from two dogs raised at farm (2.7%), and 188 worms from 26 shepherds (33.5%). No worms were recovered from the cattle or pigs. These results suggest that the dogs, especially the military dogs are serving as a reservoir host of T. callipaeda in Korea.     

Thelazia callipaeda (Spirurida, Thelaziidae) of carnivores and humans: morphological study by light and scanning electron microscopy

Thelazia nematodes cause ocular infection in several mammals and are transmitted by dipteran flies. Thelazia callipaeda Railliet and Henry 1910 (Spirurida, Thelaziidae) is common in the Far East, where it causes eyeworm infection in carnivores and humans. In the past few years, it has also been reported in dogs, cats and foxes in Northern and Southern Italy. Although a great deal of work has been carried out in China and other Eastern countries, knowledge of the morphology of T. callipaeda is scanty. Eighty-three nematodes collected from the eyes of naturally infected dogs from the Basilicata region (southern Italy) were examined by light and scanning electron microscopy, and the most important features of the nematodes were described. The morphological characteristics useful for the identification of T. callipaeda are discussed.     

Parasites of the digestive tract in beef cattle imported from France to Italy

Beef cattle heads (195 heads, 6 batches) imported for fattening from France to Italy were examined. Coprological qualitative and quantitative tests were performed, and the results analysed in relation to sex, breed, age, date of arrival, geographical origin (French department in which the animal was bred), and gathering centre (French department in which the animal was grouped with the others before travelling to Italy). The following parasites were identified: Eimeriidae (overall prevalence 60.5%); Strongyles (59%); Nematodirus spp. (14.3%); Trichuris spp. (4.1%); Capillaria spp. (2.0%); Paramphistomum spp. (27.6%); Dicrocoelium spp. (3.0%); Moniezia spp. (8.7%). All the observed parasites are widely reported in beef cattle either in Italy or in France. However, the seeming absence of Fasciola hepatica was unexpected, as well as the high prevalence of Paramphistomum spp. The variables that appeared to be more linked to parasite epidemiology were sex, altitude of the geographical origin and season.     

Thelazia rhodesii (Desmarest, 1828) in cattle of Central Province, Zambia

Sanga cattle from Central Zambia were examined during the dry and the rainy seasons for the presence of the adult stage of eyeworm Thelazia rhodesii. Prevalences were recorded according to age/sex groups. The occurrence of ocular diseases was also noted. Differences between the total prevalences of infection with the parasite in the dry and rainy seasons were observed: respectively 3.1% and 26.6% of all cattle examined (p less than 0.001), which numbered 223 head in the dry season and 248 in the rainy season. In both seasons, adult males showed the highest prevalence and calves the lowest. In the dry season the parasite was found in two of the 23 males (8.7%) but in none of the 24 calves. During the rains, eight of the 22 adult males (36.4%), and two of 53 calves (3.8%) were parasitized (p less than 0.001). A comparison of the prevalences in the latter season between a group of cows sprayed weekly with a synthetic pyrethroid and an untreated group showed no significant difference between the two groups (p greater than 0.1). No correlation was found between T. rhodesii infection and the occurrence of eye lesions (p greater than 0.1). It is concluded that in traditionally managed Zambian cattle the infection is seasonal, is more frequent in adult male animals and is not associated with eye infections (i.e., pink-eye).     

A comparison of sampling methods with respect to cattle-visiting Muscidae and their nematode infections

Abstract. Cattle-visiting Muscidae were sampled with a handnet from a blood-smeared Masonite panel as well as grazing heifers and dairy cows in south-central Sweden. All flies were examined in search of nematodes. Only Musca autumnalis was infected with nematodes (Parafilaria bovicola, Thelazia spp. and Hetero-tylenchus autumnalis). The only mixed infections found involved P.bovicola and Thelazia spp. Most M.autumnalis infected with P.bovicola and Thelazia spp. were found during the two first months of the grazing season, whereas the prevalence of specimens infected with H.autumnalis remained about the same throughout the sampling period. Musca autumnalis was caught in significantly smaller numbers from heifers (P< 0.05) than from panel and cows. The catches of nematode-infected M.autumnalis were similarly distributed. Catches of other cattle-visiting Muscidae did not differ significantly among the methods used. Handnet sampling off a panel is therefore considered to be a convenient and accurate method when investigating the abundance of cattle-visiting Muscidae and the prevalence of their nematode infections.     

Phortica variegata as an intermediate host of Thelazia callipaeda under natural conditions: evidence for pathogen transmission by a male arthropod vector

Knowledge about Phortica variegata (Drosophilidae, Steganinae), the intermediate host of the eyeworm Thelazia callipaeda (Spirurida, Thelaziidae), is confined to experimental studies. To investigate the role P. variegata plays in the transmission of T. callipaeda under natural conditions, the population dynamics of these flies in the natural environment and their feeding preferences (on vegetables and/or animal lachrymal secretions) were examined. From April to November 2005, a total number of 969 (557 males and 412 females) P. variegata flies were collected weekly in a region of southern Italy with a history of canine thelaziosis. The flies were identified and dissected or subjected to a PCR assay specific for a region within the ribosomal ITS-1 DNA of T. callipaeda. The zoophilic preferences of P. variegata were assessed by collecting flies around the eyes of a person or around a fruit bait. Seven hundred and twenty flies (398 males and 322 females) were dissected under a stereomicroscope; 249 flies (158 males and 91 females) that died prior to the dissection were subjected to molecular investigation. Only P. variegata males were infected with larval T. callipaeda both at dissection (six, 0.83%) and with the specific PCR (seven, 2.81%), representing a total percentage of 1.34% flies infected. Interestingly, only males were collected around the eyes, compared with a male/female ratio of 1:4 around the fruit. This survey indicated that P. variegata males act as intermediate hosts of T. callipaeda under natural conditions in Europe. Both the zoophilic behaviour of P. variegata males on lachrymal secretions and their role as vector of T. callipaeda have been discussed as they represent a peculiarity in medical and veterinary entomology. The synchrony between the fly population dynamics and the biology of the nematode in the definitive host provides an interesting model for exploring the co-evolution of Thelazia spp. with their hosts.     

Bacteria and nematodes in the conjunctiva of mule deer from Wyoming and Utah

Swabs of conjunctiva were collected from 44 live and 226 hunter-harvested mule deer (Odocoileus hemionus) from Wyoming and Utah (USA). We identified 29 gram negative and 22 gram positive bacterial taxonomic categories, but many isolates from hunter-harvested animals were environmental contaminants. Staphylococcus spp. and Micrococcus spp. were the most common gram positive bacteria isolated, and Enterobacter spp., Escherichia coli, and Pseudomonas spp. were common gram negative bacteria isolated. Thelazia californiensis were found in 15% of hunter-harvested deer in Utah in 1994 and in 8% in 1995. Nematodes were found in 40% of live deer in 1995 and 66% in 1996. Three live animals showed clinical signs of infectious keratoconjunctivitis (IKC) in 1996, but pathogenic bacteria were not isolated from these individuals. Hemolytic, non-piliated Moraxella ovis was isolated from two clinically normal live deer in 1996 and isolates were similar to those cultured from IKC cases from Wyoming and Utah.     

Concurrent infections of Fasciola, Schistosoma and Amphistomum spp. in cattle from Kafue and Zambezi river basins of Zambia

This study investigated interactions among Fasciola gigantica, Schistosoma spp. and Amphistomum spp. concurrent natural infections in Zambian cattle, based on egg and worm counts. In the abattoir 315 cattle were screened for worms of F. gigantica in the liver, Schistosoma spp. in mesenteric veins and/or Amphistomum spp. in the rumen. One hundred and thirty-three (42.2%) of the abattoir-examined cattle harboured one, two or all three trematodes. Of 133 cattle, 50 were randomly selected for worm and egg counts. The mean numbers (+/- SD) of Amphistomum, Schistosoma and Fasciola were 622.08 (+/- 97.87), 33.68 (+/- 7.44) and 19.46 (+/- 4.58), respectively. A total of 32% harboured all the three trematodes, 66% had F. gigantica and Amphistomum spp. infections, 52% had Schistosoma spp. and Amphistomum spp. infections while 32% had F. gigantica and Schistosoma infections. A positive correlation (P = 0.014) was found between F. gigantica and Amphistomum worm burdens. There were no correlations between Amphistomum and Schistosoma worm burdens and between F. gigantica and Schistosoma worm burdens. It may be concluded that there is no significant cross-protection among these trematodes in cattle in endemic areas.