Ovicidal, larvicidal and adulticidal activity of black pepper (Piper nigrum L.) essential oil and tea tree oil (Melaleuca alternifolia) against Haemonchus contortus.

Haemonchosis is a worldwide helminthic disease affecting ruminants. The anthelminthic resistance has become raised. Medicinal plants are safely used as synthetic anthelmintics. Currently, the efficacy of black pepper essential oil (BPO) and tea tree oil (TTO) were in vitro evaluated against Haemonchus contortus adults, eggs and larvae at concentrations of 1.25, 2.5 and 5 mg/ml in addition to the commercially used albendazole at a concentration of 10 µg/ml. Oils were used in both normal and nanoparticles-loaded forms. Oxidative stress enzymes of worms were estimated. Scanning electron microscopy (SEM) for treated worms was done. Both normal and nanoemulsion forms of both BPO and TTO stopped the adult motility [BPO 2.5 h (hrs), NBPO 1.5 h, TTO 3 h, NTTO 1.5 h] and induced a marked decrease in the oviposition. Post treatment, the egg development and hatching were significantly (P ≤ 0.05) reduced. The damage of the egg shell, embryonal cessation and destruction of larvae occurred. Noticeable elevated antioxidant enzymes (catalase CAT, glutathione transferase GST and glutathione GSH) were found, while oxidative enzymes (lipid peroxidation LPO and nitric oxide synthase NOS) decreased. Scanning electron microscopy (SEM) for both oil-treated worms revealed anterior ends damage and several cuts associated with cuticular pores. The use of albendazole induced more or less anthelmintic and enzymatic activities with less morphological alterations of adults revealed by SEM. This study proved the marked anthelmintic potency of the BPO and TTO and their nanoemulsion forms against H. contortus rather than the widely used anthelmintic drugs.

The prevalence and molecular detection of bovine cysticercosis and its impact on slaughtered cattle in Egypt.

Cysticercus bovis is the metacestode of the commonly defined foodborne cestode, Taenia saginata. It infects heart, masseter muscles and other muscular sites of cattle causing bovine cysticercosis. So, a retrospective study using archival documents from authorized agencies is done during the period of 2018-2020 with estimating economic losses. Cysts were collected from municipal Beni-Suef and Basatin abattoirs. Molecular approach including cPCR and sequencing analysis is used to prove the species. The prevalence was 0.34% in Beni-Suef abattoir and 5.70% in Basatin abattoir including heads, hearts or all carcasses condemned. Among 27 provinces surveyed, the highest prevalence was recorded in Aswan followed by Cairo, Suez and Red Sea. Moreover, 19 provinces (Cairo, Alexandria, Suez, Dakahlia, Sharkia, Qalyobia, Kafrelsheikh, Gharbia, Menofiya, Ismailia, Giza, Beni-Suef, Assiut, Sohag, Qena, Aswan, Red Sea, South Sinai and El-Wadi El-Gadid) recorded bovine cysticercosis in 2018, 2019 and 2020. On the other hand, Luxor and North Sinai had no infections. The percentages of condemnation among slaughtered cattle were 1.38, 1.49 and 0.87% in 2018, 2019 and 2020, respectively. Condemnations significantly varied among north, middle and south districts of Egypt, and also varied annually. Molecularly, a diagnostic band at an amplicon size 253 bp targeting the COI gene specific for Cycticercus bovis was revealed. The obtained sequences showed 100% identity with the different Taenia saginata COI GB sequence isolates in many countries worldwide. The phylogenetic analysis method showed that the obtained sequences originating from the same clade of Taenia saginata GB isolates globally. Careful meat inspection as well as strict hygienic measures is recommended for both veterinarians and public.

Economic losses, morpho-molecular identification, and identity of Fasciola species recovered from Egypt.

A retrospective study to estimate economic losses caused by livers condemnation, due to fascioliasis, of slaughtered cattle and buffaloes in Egypt during the period of 2016-2020, was done. Moreover, a morpho-molecular identification of collected liver flukes from slaughtered animals in municipal abattoirs was conducted. Livers of naturally infected carcasses were obtained from slaughtered animals in Beni-Suef, Cairo and Tanta provinces, Egypt during 2019-2020 for phenotypic characterization of recovered Fasciola species and molecular identification of collected worms using PCR targeting the ITS-1 region. Findings of the retrospective study revealed that percentages of livers condemnation of cattle and buffaloes ranged from 0.79 to 0.66% during the period from 2016 to 2020. The highest percentages were detected in the south Egypt (2.5-6.0%) with the highest economic losses (261850-616300 USD annually). Morphometrically, collected flukes categorized into Fasciola hepatica and Fasciola gigantica. No intermediate forms (Fasciola sp.-like) were detected. Sequencing analysis of ITS-1 PCR products showed that only Fasciola hepatica (26/34) and F. gigantica (8/34) isolates were found, with no intermediate forms, Fasciola sp.-like, could be identified. Currently, Fasciola hepatica was 100% identical with the Egyptian species (LC076196 and JF294998), French species (JF294999), and Iranian species (MF969009 and MK377150). Moreover, the obtained F. gigantica species showed 100% identity with Egyptian ones (LC076125, LC076108 and KX198619), Iranian (KF982047 and MF372919), and other GenBank specimens from Vietnam, Cameroon and India. In conclusion, South Egypt showed the highest economic losses due to fascioliasis, especially Aswan province. Fasciola hepatica was more common than F. gigantica, while the hybrid form was not detected.

Occurrence of Dirofilaria immitis in Stray Dogs from Nepal.

BACKGROUND: Nematodes of the genus Dirofilaria are widespread vector-borne helminths (VBH) of veterinary relevance. Dirofilariosis caused by D. immitis is frequently diagnosed in dogs often showing a severe clinical condition known as heartworm disease. Assessing the distribution pattern of canine dirofilariosis is pivotal to undertake appropriate control measures and define the risk of infection in animals and humans. This study provides original data on the occurrence of D. immitis in naive stray dogs from Nepal. METHODS: An epidemiological study was performed on stray dogs from Siddharthanagar, Lumbini region, Nepal. Alive animals were blood sampled and examined for microfilariae using direct microscopy, buffy coat centrifugation and modified Knott's method. In addition, hematobiochemical parameters, including packed cell volume (PCV), alanine aminotransferase (SGPT), aspartate aminotransferase (SGOT), alkaline phosphate (ALP), creatinine, blood urea nitrogen (BUN), urea and total protein were analysed, along with a clinical scoring (body temperature and respiratory rate). During the observation period, data on autopsies performed on stray dogs showing canine heartworm disease were also included. RESULTS: The overall prevalence of D. immitis in both alive and necropsied dogs was 22.49% (38/169). Out of 150 alive dogs, 29 (19.3%) were positive for D. immitis microfilariae. The prevalence of the infection varied according to the technique used, ranging from 16.0 to 19.3% based on direct blood smear microscopy and modified Knott's technique, respectively. The infection rate was significantly higher (p < 0.05) in dogs aging more than 3 years. A positive correlation was observed between SGPT and SGOT and the presence of microfilariae (p < 0.05). Among the 19 dead stray dogs, 9 animals showed adult D. immitis nematodes. CONCLUSION: Results of this survey account for the presence of D. immitis in stray dogs from Nepal, thus supporting the existence of a local transmission cycle of diroflariosis in the region. Further investigations are warranted to accurately define the prevalence of the infection in the canine population and to undertake appropriate control measures to reduce the burden on the infection in dogs and, accordingly, the risk of transmission to humans.

Molecular Detection, Phylogenetic Analysis, and Genetic Diversity of Theileria annulata, Babesia bigemina, and Anaplasma marginale in Cattle in Three Districts of Egypt.

BACKGROUND: Under the poor hygienic conditions, tick-borne pathogens cause severe economic losses to the cattle industry. PURPOSE: The current study investigated the presence of Theileria annulata, Babesia bigemina, and Anaplasma marginale, the most relevant tick-borne pathogens in cattle, in 3 provinces of Egypt utilizing species-specific PCR assays. METHODS: PCR was conducted, on bovine blood specimens, using primers targeting the T. annulata merozoite-piroplasm surface antigen (Tams1, 768 bp), A. marginale major surface protein-1b gene (msp1b, 265 bp), and B. bigemina small subunit ribosomal RNA gene (SSrRNA, 543 bp). RESULTS: PCR findings revealed overall prevalences of T. annulata, B. bigemina, and A. marginale as 22.0% (33/150), 19.33% (29/150), and 10.6% (16/150), respectively. The co-infection with two or three pathogens was detected in 20.0% (30/150) of examined specimens. Sequence analyses indicated that T. annulata and A. marginale varied from those of corresponding GenBank sequences revealing percent identities ranging from 90.68 to 97.75% and from 94.98 to 98.63%, respectively. On the other hand, the obtained B. bigemina sequences showed a high similarity with those previously reported in GenBank with a percent identity ranging from 98.85 to 100%. CONCLUSION: T. annulata was the most prevalent tick-borne pathogen in examined bovine specimens. The genetic diversity of markers used for identification of T. annulata and A. marginale should be highly considered.

The current prevalence and diversity of cystic echinococcosis in slaughtered animals in Egypt.

Cystic echinococcosis is a potential zoonotic helminthic disease affect a broad spectrum of mammals including humans worldwide. The economic impact of the disease interestingly requires updated gathering information on the prevalence in slaughtered animals. Accordingly, in the current study, 573 camels, 4300 sheep, and 1235 pigs were surveyed in four Egyptian municipal abattoirs. Among those, 62 (10.82%) camels, 33 (0.77%) sheep and 3 (0.24%) pigs had cystic echinococcosis in lungs, livers and spleen. The diversity of cysts revealed that positive-cystic echinococcosis animals from all species were detected in El-Basatin abattoir. In El-Monieb abattoir, sheep only were infected. In El-Waraa and Beni-Suef abattoirs, cysts were seen in camels only. Infected animals included both sexes. In camels, lungs were the most affected organs, while in sheep, livers were the most abundant. In pigs, hydatid cysts were detected only in lungs. Moreover, camel cysts were mostly large-sized (diameter > 10 cm) with smaller cysts are also recorded. In sheep, small-, moderately- and large-sized cysts were equally present. Cysts in pigs were large-sized. Camels showed a higher percentage of fertile cysts (46.77%; 29/62) followed by sheep (21.21%; 7/33). Calcified/degenerated cysts were less in camels (38.71%; 24/62) than in sheep (75.76%). Non-viable cysts were the less frequent in both species (14.52% in camels and 3.03% in sheep). All cysts recovered from pigs were fertile. In Egypt, governmental agencies and veterinary authorities are asked to potentially eradicate stray dogs (the main definitive host) and towards the proper hygienic disposal of infected offal in abattoirs to minimize the prevalence of cystic echinococcosis.

Prevalence and distribution pattern of intestinal helminths in chicken and pigeons in Aswan, Upper Egypt.

There is scarce data available on helminth infection of poultry in the region of Aswan province, Upper Egypt. A total of 276 poultry (156 baladi chickens and 120 domestic pigeons) were collected from different villages in Aswan province to screen the presence of intestinal helminths during the period from April 2016 to March 2017. The current investigation revealed that the overall prevalence was 55.79% (154/276). Additionally, 59.09% baladi chickens had mixed infections with four cestodes and three nematodes. Among cestodes, Raillietina tetragona was the most prevalent species (16.02%) followed by Raillietina echinobothrida (10.98%), Cotugnia digonopora (6.41%), and Raillietina cesticillus (1.28%). The most prevalent nematodal species were Heterakis gallinarum (15.38%), Ascaridia galli (8.97%), and Subulura brumpti (1.28%). In pigeons, 52.5% were found to be infected by four cestodes: Raillietina echinobothrida (22.5%), Cotugnia digonopora (13.33%), Raillietina tetragona (9.16%), and Raillietina cesticillus (3.33%) as well as one species of nematodes, Ascaridia columbae (9.16%). There was a close relationship between the occurrence of intestinal helminths and the age of domestic birds revealing that the high prevalence was found in adults rather than young birds. Meanwhile, it has been found that the highest prevalence was found in summer, while the lowest infection rate was recorded in winter. Further studies are needed to determine the effects of the parasites on the health and productivity of poultry in such area.

A case report on the gallbladder fluke, Euparadistomum herpestesi (Trematoda: Dicrocoeliidae), in stray cats (Felis catus) in Beni-Suef, Egypt.

Scarce literature studying parasites, particularly helminths, of gallbladder of cats in Egypt are available. Stray cats (Felis catus) are widespread in Beni-Suef, Egypt. Therefore, a total of 62 stray cats were necropsied and parasitologically examined. Three (4.84%) cats harboured an ovoid yellowish to brown flukes in the gallbladder. Infected cats were females and aged more than 3 years. The worm burden ranged 1-3 (one cat harboured 3 flukes and the other 2 cats were infected by one fluke each). Moreover, Mixed infections with intestinal cestodes and/or nematodes were detected. The fluke is quite discoid measuring 7.75 × 3.25 mm. The oral sucker measured 1 mm in diameter. The acetabulum was nearly central, rounded and measured 0.75 mm in diameter. Short pharynx (no prepharynx). Two broad intestinal ceca extending short distance from the posterior extremity of the fluke. Testes are ovoid, opposite to each other occupying the anterior half, each measured 0.25 mm in diameter. The ratio of testes to the acetabulum is characteristically 1:2. Ovary was somewhat ovoid, measured 0.3 × 0.45 mm, and attached to it a large dorsal chamber of receptaculum seminis measuring 0.58 × 0.52 mm. Vitellaria extended from the level of testes along the length of intestinal caeca or somewhat shorter than length of them. Uterus consisted of several coils occupying the entire area behind the oral sucker. Based on those criteria, the recovered flukes might be identified as Euparadistomum herpestesi. To the best of authors' knowledge and based on the microscopic identification, it is the first preliminary report denoting the existence of the gallbladder flukes, E. herpestesi, from stray cats in Beni-Suef, Egypt.

Ecological study on antimicrobial-resistant zoonotic bacteria transmitted by flies in cattle farms.

Flies were qualitatively and quantitatively monitored on both livestock animals and the surrounding environment to investigate their role as a potential carrier for antimicrobial-resistant bacteria of zoonotic importance in cattle farms. This was done by the use of visual observations and animal photography; meanwhile, in the surrounding environment, flies were collected using sticky cards and then microscopically identified. Representative fly samples were cultured for bacterial isolation, biochemical identification, and then tested against common 12 antibiotics. The total average of dipterous flies in examined farms was 400.42 ± 6.2. Culicoides biting midges were the most common existing species (70.01 %) followed by house flies, stable flies, and mosquitoes (18.31, 7.74, and 3.91 %, respectively) at X (2) = 9.0, P < 0.05. The most predominant bacterial isolates were Escherichia coli (22.6 %), Staphylococcus aureus and Enterobacter (17.3 % each), coagulase-negative Staphylococci (CNS) (14.7 %), Klebsiella sp. (8 %), Salmonella spp. (6.7 %), and Shigella spp. and Proteus spp. (6.7 % each). The tested bacterial isolates were resistant to variant antibiotics used. S. aureus exhibited 100 % resistance to colistine. However, E. coli revealed 92.9 and 78.6 % resistance against tetracycline and colistine, respectively. Both Salmonella spp. and Shigella spp. were 100 % resistant to penicillin, and Klebsiella sp. had 100 % resistance to tetracycline. In conclusion, Culicoides biting midges and house flies could be considered as a potential carrier for multi-drug-resistant bacteria of zoonotic importance. Furthermore, cows' environment has an essential role in propagation and wide spread of antimicrobial-resistant bacterial pathogens.

Morphologic and molecular characteristics of Sarcocystis atraii n. sp. (Apicomplexa: Sarcocystidae) infecting the common coot (Fulica atra) from Egypt.

A single morphologic type of Sarcocystis cysts found in two out of 43 examined common coots, Fulica atra, is considered to represent a new species for which the name Sarcocystis atraii n. sp. is proposed and its description is provided. Coots were hunted from the vicinity of Brolos Lake located at KafrElsheikh province, Egypt. The structural morphology of the revealed sarcocysts was described using light and transmission electron microscopy. Sarcocysts were found in the leg and thigh muscles. The cysts were microscopic and measured 165-850 µm in length × 50-85 µm in width. Histologically; the sarcocyst wall was wavy and had minute undulations. Ultrastructurally, it measured 1-3 µm in thickness and possessed many mushroom-like villar protrusions sometimes originating from other mushroom-like villar protrusions that measured approximately 0.5-2 µm in length and up to 2 µm in width, with the presence of electron dense ground substance of 300 nm to 1 µm thick. The bradyzoites were elongated, banana-shaped and measured 7.5-14 × 1.5-2.5 µm, with centrally or terminally located nuclei. The ultrastructural features of the cyst wall belonged to type 24. On the basis of sequencing and phylogenic analyses for 18S rRNA , 28S rRNA genes and ITS-1 region; S. atraii n. sp. is considered a genetically distinct species, being most closely related to avian Sarcocystis spp. whose definitive hosts are predatory mammals.

Distribution patterns of Babesia gibsoni infection in hunting dogs from nine Japanese islands.

Canine babesiosis constitutes a major global veterinary medical problem caused by tick-borne hemoparasites Babesia gibsoni and Babesia canis. Babesia gibsoni induces more severe clinical signs and is mainly transmitted by the ixodid Haemaphysalis longicornis. In Japan, B. gibsoni is primarily found in the western districts, with few records in the eastern parts. The aim of the current investigation was to evaluate distribution patterns of B. gibsoni infection in 9 Japanese islands and peninsulas using direct microscopy and PCR. Therefore, 196 hunting dogs were randomly sampled during the period from March to September 2011. Ages and sexes of dogs were identified. Direct microscopy of Giemsa-stained blood smear revealed pear-shaped piroplasms of B. gibsoni in 3 (1.6%) dogs. PCR was done initially with the universal primer set (B18S-F and B18S-R) amplifying the 1,665-bp portion of the 18S rRNA gene, followed by the specific primer set (Bg18F1 and Bg18R2) amplifying 2,363-bp fragments of the same gene. Accordingly, 84 (42.9%) and 8 (4.1%) dogs were positive, respectively. The current investigation shows that canine babesiosis was recorded in all islands except for Sado Island, Atsumi Peninsula, and Tanegashima Island. The highest infection rate was detected in the main island of Okinawa, while the lowest was on Ishigaki Island. Both sexes were non-significantly infected. However, the diversity of infection in islands was significantly different (P < 0.05). Although B. gibsoni has been previously found in western and eastern Japan, the present work highlights the prevalence of infection in many Japanese districts, including islands and peninsulas, giving realistic data that can facilitate treatment and control.

Monitoring of parasitic cysts in the brains of a flock of sheep in Egypt.

Cerebral parasitic cysts constitute a major problem for livestock. Among these, coenurosis and toxoplasmosis are predominant. Here, a total number of 60 sheep obtained from a private farm in Suez province, Egypt, were examined postmortem to detect visible parasitic cysts, and microscopically to detect small-sized entities. Necropsy revealed bladder-like cysts measuring 0.5-6.5 cm in diameter that were filled with a translucent fluid containing a large number of protoscolices. Accordingly, the cysts were identified as the metacestode Coenurus cerebralis. Among the sheep examined, 11 animals (7 males and 4 females) (18.3%) were infected. Most of the cysts were located in the cerebral hemispheres, with numbers ranging from one to three per infected animal. The effect of the presence of cysts in the brain tissue was evaluated. Histopathologically, pseudocysts of the apicomplexan Toxoplasma gondii were found in two animals with no detectable inflammatory cell reactions. In conclusion, coenurosis and toxoplasmosis are serious parasitic problems that play a significant role in sheep management in Egypt, as a result of close contact between livestock and dogs and cats, which play a critical role in the life cycle of these parasites.

Monitoring of parasitic cysts in the brains of a flock of sheep in Egypt / Monitoramento de cistos parasitários no cérebro de um rebanho de ovinos no Egito

Cerebral parasitic cysts constitute a major problem for livestock. Among these, coenurosis and toxoplasmosis are predominant. Here, a total number of 60 sheep obtained from a private farm in Suez province, Egypt, were examined postmortem to detect visible parasitic cysts, and microscopically to detect small-sized entities. Necropsy revealed bladder-like cysts measuring 0.5-6.5 cm in diameter that were filled with a translucent fluid containing a large number of protoscolices. Accordingly, the cysts were identified as the metacestode Coenurus cerebralis. Among the sheep examined, 11 animals (7 males and 4 females) (18.3%) were infected. Most of the cysts were located in the cerebral hemispheres, with numbers ranging from one to three per infected animal. The effect of the presence of cysts in the brain tissue was evaluated. Histopathologically, pseudocysts of the apicomplexan Toxoplasma gondii were found in two animals with no detectable inflammatory cell reactions. In conclusion, coenurosis and toxoplasmosis are serious parasitic problems that play a significant role in sheep management in Egypt, as a result of close contact between livestock and dogs and cats, which play a critical role in the life cycle of these parasites.

Cistos cerebrais parasitários constituem um grande problema para o gado. Entre estes, coenurosis e toxoplasmose são predominantes. Aqui, um número total de 60 ovelhas obtidas em uma fazenda particular na província Suez, Egito, foram examinadas post-mortem para a detecção de cistos parasitários visíveis e microscopicamente para detectar cistos de pequenas dimensões. A necropsia revelou cistos medindo entre 0,5-6,5 cm de diâmetro, preenchidos com um fluido transparente, contendo um grande número de protoscolices. Por conseguinte, os cistos foram identificados como o metacestóide Coenurus cerebralis. Entre as ovelhas examinadas, 11 animais (7 machos e 4 fêmeas) (18,3%) estavam infectados. A maior parte dos cistos estavam localizados nos hemisférios cerebrais, com números variando de um a três em ovinos infectados. O efeito da presença de cistos no tecido do cérebro foi avaliado. Histopatologicamente, pseudocistos de Toxoplasma gondii foram encontrados em dois animais sem reações inflamatórias detectáveis. Em conclusão, coenurosis e toxoplasmose são graves problemas parasitários que desempenham um papel significativo no manejo de ovelhas no Egito, como resultado do contacto íntimo dos animais com os cães e gatos, que desempenham um papel crítico no ciclo de vida desses parasitas.

Prevalence and diversity of Hepatozoon canis in naturally infected dogs in Japanese islands and peninsulas.

Canine hepatozoonosis is a worldwide protozoal disease caused by Hepatozoon canis and Hepatozoon americanum and is transmitted by ixodid ticks, Rhipicephalus and Amblyomma spp., respectively. H. canis infection is widespread in Africa, Europe, South America, and Asia, including Japan. The objective of this study was to study the distribution pattern and diversity of H. canis in naturally infected dogs in nine Japanese islands and peninsulas. Therefore, 196 hunting dogs were randomly sampled during the period from March to September 2011 and the ages and sexes were identified. Direct microscopy using Giemsa-stained blood smears revealed H. canis gametocytes in the peripheral blood of 45 (23.6%) dogs. Polymerase chain reaction (PCR) was performed on EDTA-anticoagulated blood, initially with the common primer set (B18S-F and B18S-R) amplifying the 1,665-bp portion of the 18S rRNA gene, and then with the specific primer set (HepF and HepR) amplifying about 660 bp fragments of the same gene. Based on PCR, 84 (42.9%) dogs were positive using the common primer and 81 (41.3%) were positive using the specific primer. The current investigation indicated that all screened areas, except for Sado Island and Atsumi Peninsula, were infected. Yaku Island had the highest infection rate (84.6% in males and 100.0% in females), while Ishigaki Island showed the lowest infection rates (8.3% in males and 17.7% in females). Both sexes were infected with no significant difference. However, diversity of infection among the surveyed islands and peninsulas was significantly different (P < 0.05). Although H. canis has previously been reported in dogs in Japan, the higher infection rate described in the current study and the diversity of infection in a wide range of islands strongly encourage prospective studies dealing with the prevention and treatment of the infection in dogs, as well as control of ticks.

Hypodectes propus (Acarina: Hypoderatidae) in a rufous turtle dove, Streptopelia orientalis (Aves: Columbiformes), in Japan.

An adult male rufous turtle dove, Streptopelia (S.) orientalis (Aves: Columbiformes), was found dead in Yorii-machi Town, Osato District 369-1217, Saitama Prefecture, Japan, and subjected to necropsy. A large number of immobile hypopi (deutonymphs) of the hypoderatid mite, Hypodectes (H.) propus (Acarina: Hypoderatidae), were found individually encapsulated subcutaneously primarily in the adipose tissue. The mites were 1.43 mm in length and 0.44 mm in width on average, and had provoked mild inflammatory reactions that predominantly manifested as foamy macrophages and lymphoplasmocytes. PCR analysis using ribosomal DNA extracted from paraffin-blocked tissues produced a 240 bp band specific for hypoderatids. Based on the morphological features (distinct coxal apodemes, especially in the anterior portion) and PCR-based findings, the hypopi were identified as H. propus. To the best of our knowledge, this is the first case describing the subcutaneous mite H. propus in a rufous turtle dove, S. orientalis, in Japan. This study also highlights the use of paraffin blocks as a source of tissue DNA for molecular evaluation.

Molecular characterization of Echinococcus granulosus in Egyptian donkeys.

The present study was carried out during the period from August 2009 to July 2010. The purpose of this study was to identify the genotype of donkeys' echinococcosis in one of the Egyptian governorates. On post-mortem inspection taking place at the zoo of Beni-Suef, Egypt, a total number of 145 donkeys were examined for the presence of hydatid cysts. Ten of these donkeys were found to be infected by hydatid cysts; and location, number and fertility of cysts found were determined. The liver was the predominant site of infection. Molecular identification of these cystic echinococcosis isolates, based on PCR amplification and the sequence of both mitochondrial ND1 and CO1, revealed that they belonged to Echinococcus equinus (G4 genotype). An alignment of ND1 and CO1 partial nucleotide sequences with G4 partial nucleotide sequences revealed replacement of G at position 105 with A and replacement of A at position 276 with G respectively. It can be concluded that the donkeys involved in this study were harboring E. equinus. For the first time in Egypt, the present work allowed us to record the presence of the E. equinus with the molecular tools, and to report new information on the epidemiological status of this parasite in Egypt.

The use of different diagnostic tools for Babesia and Theileria parasites in cattle in Menofia, Egypt.

Bovine piroplasmosis is caused by tick-borne hemoprotozoans of the genera Babesia and Theileria and is the most prevalent in tropical and subtropical countries, causing a major economic impact worldwide. In the current study, a total of 405 cattle of different ages, sexes, and breeds were randomly sampled for surveying and diagnosis of babesiosis and theileriosis using three methods: direct microscopy (blood smears), indirect fluorescent antibody test (IFAT) and polymerase chain reaction (PCR). Giemsa-stained blood smears revealed that, out of 405 examined cattle, 33 (8.15 %) were infected with Babesia sp. and 65 (16.05 %) with Theileria sp. (total number of infected cattle was 98). Mixed infection was seen in 11 (2.72 %) animals. Moreover, application of the three diagnostic assays on 158 randomly sampled cattle indicated that 17 (10.76 %) and 33 (20.89 %) were positive for Babesia and Theileria spp. by the direct smear technique, 25 (15.82 %) and 33 (20.89 %) by IFAT (fluorescence was greenish yellow for Babesia and yellowish for Theileria), and 20 (12.66 %) and 38 (24.05 %) by PCR. Using primers specific for Babesia and Theileria spp., we found that diagnostic bands appeared at ~350 and ~370 bp, respectively indicating the presence of these piroplasms. Statistically, there was a non-significant difference of the positivity in response to the three techniques; thus, any of these methods can be described as useful for diagnosing blood parasites in both domesticated animals and birds. On the basis of the obtained results, it could be concluded that direct microscopy can be used in acute infections, whereas IFAT and PCR are useful in chronicity.

Prevalence and distribution patterns of Sarcocystis spp. in buffaloes in Beni-Suef, Egypt.

This study was performed for the purpose of investigating the prevalence of Sarcocystis spp. in buffaloes in Beni-Suef Governorate, Egypt. Both macroscopic (Sarcocystis fusiformis) and microscopic (Sarcocystis levinei) cysts were recognized, and were differentiated by their morphological features and location in the tissues. Of 379 buffaloes examined in abattoirs in Beni-Suef, 299 were found to be infected, with an overall prevalence of 78.9%. Depending on age, three categorized groups of naturally infected buffaloes were examined: male buffalo calves aged 1.5-2 years, adult females aged 2-5 years, and females older than 5 years. Among these groups, infection rates were 74.5%, 82.3%, and 81.2%, respectively. Organs examined included esophagus, tongue, and heart. Macroscopic cysts were examined by the naked eye through meat inspection in abattoirs, while the pepsin-digestion method and the histological technique were applied to detect microscopic cysts. It has been found that esophagus showed the highest rate of infection among the infected organs, with both macroscopic and microscopic cysts seen in the infected buffaloes. Moreover, results of the pepsin-digestion method proved more accurate than those produced by the histological technique in terms of infection rates for the microscopic cysts. Our findings indicated that infected buffaloes aged 2-5 years showed the highest mixed infection rate (82.3%) for both types of cysts. The high prevalence of microscopic Sarcocystis spp. in Beni-Suef Governorate reflects a significant role played by stray dogs, rather than cats, in the transmission of these parasites.