Prevalence and molecular identification of Nematodirus helvetianus in camels in Iraq.

BACKGROUND AND AIM: Camels from the central part of Iraq are infected with multiple parasitic diseases that have an economic impact by decreasing meat and milk production. This study aimed to evaluate Nematodirus spp. in camels (Camelus dromedarius). MATERIALS AND METHODS: The study animals consisted of camels slaughtered in the central area of Iraq at the Al-Najaf slaughterhouse. All ages and sexes of camels were examined. Worms were recovered and identified microscopically. For molecular characterization, two Iraqi Nematodirus spp. partial ribosomal genes (ITS1 and ITS2) were sequenced and submitted to the NCBI database. RESULTS: Of 160 camels tested, 29 were infected with Nematodirus spp. (18.13%). Twenty-one nematodes containing the Nematodirus genes were identified in the small intestines of naturally infected camels. BLAST analysis revealed 88.1% sequence similarity with that of Nematodirus helvetianus isolated in China and 87.2% similarity with N. helvetianus isolated in the United States. CONCLUSION: The prevalence of N. helvetianus warrants the use of anti-helminthic drugs for these animals and a rationale for future control strategies to prevent the transmission of this infection to other livestock.

In vitro cultivation of Babesia duncani (Apicomplexa: Babesiidae), a zoonotic hemoprotozoan, using infected blood from Syrian hamsters (Mesocricetus auratus).

Human babesiosis, a tick-borne disease similar to malaria, is most often caused by the hemoprotozoans Babesia divergens in Europe, and Babesia microti and Babesia duncani in North America. Babesia microti is the best documented and causes more cases of human babesiosis annually than all other agents combined. Although the agents that cause human babesiosis are considered high-risk pathogens in transfusion medicine, federally licensed diagnostics are lacking for B. duncani in both the USA and Canada. Thus, there has been a need to develop and validate diagnostics specifically for this pathogen. In this study, B. duncani (WA1 isolate) was cultivated in vitro from Syrian hamster (Mesocricetus auratus) infected blood. We hypothesized HL-1 media with supplements would result in B. duncani propagating at higher levels in culture than supplemented M199 similar to the medium the parasite was originally cultivated with in 1994. We were unable to recreate Thomford's cultivation results with the M199 medium but supplemented HL-1 medium was able to successfully establish continuous culture. We further hypothesized that RBC from species other than hamsters would support B. duncani in vitro. However, rat, mouse, horse, and cow RBC did not support continuous culture of the parasite. Culture stocks of B. duncani were deposited at BEI Resources and are now commercially available to the scientific community to further research. The cultured parasite developed in this study was instrumental in the adaptation of B. duncani continuous culture to human RBC.

Recovery of bovine Babesia spp. after long-term cryostorage and comparison of bovine donor erythrocytes and serum.

Cultured Babesia bovis and Babesia bigemina were recovered from liquid nitrogen storage nearly 30 years after they were cryopreserved. Four cattle were compared as donors of erythrocytes and serum for microaerophilous stationary phase (MASP) cultures for recovery of B. bigemina. Erythrocytes and serum from only one (#913) of the four animals supported growth of B. bigemina. Two B. bigemina (frozen in 1986 and 1987) and two B. bovis (both frozen in 1986) cryostocks were recovered from liquid nitrogen storage and all four recovered and thrived in #913 erythrocytes and serum. In the third passage after recovery, B. bovis cultures were cryopreserved. Six months later they were successfully recovered using #913 erythrocytes and serum. This study shows that B. bovis and B. bigemina stored nearly 30 years in liquid nitrogen can be successfully recovered in the MASP system. This study also confirms previous observations that selection of a suitable bovine donor of erythrocytes and serum is critical to the success of the culture.

Molecular and morphological characterization of a haemogregarine in the alligator snapping turtle, Macrochelys temminckii (Testudines: Chelydridae).

A severely underweight alligator snapping turtle Macrochelys temminckii Troost in Harlan, 1835, was found near Tyler, Texas, and taken to the Caldwell Zoo. Blood films were submitted to Texas A&M University, College Station, Texas, for morphological and molecular identification of haemogregarine-like inclusions in the red blood cells. Intraerythrocytic Haemogregarina sp. forms were found on microscopic examination at a parasitemia of <1 %. The morphology and morphometric data for the forms indicate similarity to Haemogregarina macrochelysi n. sp. Telford et al., 2009, previously reported in alligator snapping turtles in Florida and Georgia, but two characteristic stage forms were not shared between H. macrochelysi n. sp. and the parasite found in this report. The haemogregarine 18S ribosomal RNA gene (1555-bp fragment) was amplified and cloned, and five clones sequenced. The sequences were deposited in the NCBI GenBank database. All five showed ∼96 % identity to Haemogregarina balli Paterson and Desser, 1976, Hepatozoon sp., and Hemolivia stellata Petit et al., 1990. A 774-bp segment shared 98-99 % identity with the corresponding Haemogregarina sp. rDNA sequence (KR006985) from Caspian turtles (Mauremys caspica McDowell, 1964) in Iran. A neighbor-joining phylogenetic tree generated from aligned sequences from the clones, 26 hematozoa, Adelina dimidiata Schneider, 1875, and Cryptosporidium serpentis Levine, 1980, revealed the cloned sequences clustered on their own branch within the Haemogregarina spp. clade. No genetic data are available for H. macrochelysi n. sp. at this time, so it remains unclear if this parasite in a Texas alligator snapping turtle is conspecific with H. macrochelysi n. sp.