Drop-off rhythms and survival of Hyalomma anatolicum (Acari: Ixodidae) fed on crossbred (Friesians x Zebu) calves in Sudan.

Hyalomma anatolicum is one of the most economically important ticks in the Sudan. It is the main vector of tropical theileriosis in cattle and other diseases in different species. The study aims to investigate the drop-off rhythms and survival periods of tick stages fed on crossbred calves. Larvae, nymphs and adults fed on crossbred calves (male, 3-6 months old, Friesians x Zebu) kept under zero grazing system. Drop-off rhythms of engorged stages were studied under field conditions while the survival periods of unfed stages were investigated under field and laboratory conditions (27 °C and 85% R.H.). Significantly high numbers of engorged larvae dropped at night between 20H:00 and 07H:00. (March, April and August). Engorged nymphs dropped late evening and early night 16H:00-20H:00. (September), while insignificant number of engorged females dropped between 10H:00 and 14H:00 (October). Survival durations of unfed H. anatolicum stages were longer under laboratory conditions than in the field. These were 99.36 ± 1.24 and 13.12 ± 0.68 days for larvae, 63 ± 1.33 and 16 ± 0.87 days for nymphs and 90 ± 3.6 and 45 ± 2.7 days for adults, respectively. Under field condition survival studies were conducted in May, August and December 2016 for larvae, March, May, August and December 2016 for nymphs and in October 2016 for Adults. In August, 70% of unfed larvae and nymphs survived for three weeks and 2.5 weeks, respectively, in May, 16% of unfed larvae survived for one week while no unfed nymphs survived for more than 4 days. This finding may indicate that larvae are more resistant to desiccation than nymphs. The abnormal changes on the alloscutum of H. anatolicum desiccated nymphs and females were observed being shrunken and truncated posteriorly, a finding which opens new avenues for further studies. For control strategies more studies are recommended on the dropping to allow ticks drop in an area not suitable for further development as one of the control strategies and the survival studies also is exploited in rotational grazing of animals.

The pulmonary involvement in Theileria lestoquardi naturally infected sheep.

Malignant Ovine Theileriosis (MOT) caused by Theileria lestoquardi is considered a major constraint for sheep production in many areas of the world including Sudan. Pulmonary oedema is thought to be the main cause of animal death, but the mechanism, the cell types involved and/or the probable cause of this pneumonia has yet to be defined. The present study was carried out to investigate the pulmonary involvement post T. lestoquardi infection and to identify the cell types involved in pneumonia. Apparently healthy sheep were exposed to ticks challenge in T. lestoquardi endemic area. Lungs impression smears and tissue sections for histopathology were processed. At necropsy, fifteen infected sheep revealed severe pneumonia associated with oedema and accumulation of creamy-grayish frothy exudates. The microscopic findings of examined lungs showed emphysema, congestion, collapse and proliferation of immense amount of different kinds of cells. The current study indicates that T. lestoquardi infections are accompanied with remarkable pulmonary involvements and may lead to respiratory failure and death.

Molecular identification of different Theileria and Babesia species infecting sheep in Sudan.

The epidemiological aspects of sheep piroplasmosis in Sudan are poorly studied, and further investigations using sensitive and precise techniques are required. In this study, the Reverse Line Blot (RLB) hybridization assay was used to detect and simultaneously differentiate between Theileria and Babesia species. DNA was extracted from blood collected on filter paper (n=219) from apparently healthy sheep from six different geographical localities in Sudan. Results indicated that Theileria ovis (88.6%), T. separata (20.1%), T. lestoquardi (16.4%) and T. annulata (16.4%) DNA could be detected in the blood samples. Single and mixed Theileria infections were detected in 74 (33.8%) and 124 (56.6%) respectively and T. ovis being the most prevalent species in the country. T. ovis and T. separata were reported for the first time in sheep in Sudan.

Current situation of tropical theileriosis in the Sudan.

Tropical theileriosis has long been recognized as a hindrance to the development of sound dairy industry in the Sudan and is a cause of major economic losses. Serological surveys indicated that Theileria annulata infection is widespread in the country but the disease mostly affects exotic dairy breeds and their crosses with indigenous breeds. The disease has recently been identified in Darfur and southern parts of Blue Nile State in dairy farms around large urban areas where it has never been detected before. These new introductions were accompanied by the establishment of Hyalomma anatolicum tick which is the main vector of the parasite in the Sudan. The disease is routinely diagnosed using microscopic examination of stained blood and lymph node biopsy smears. More advanced techniques are mainly used for research purposes. Tropical theileriosis in the Sudan is mainly controlled by using anti-theilerial drugs and acaricide application. It is recommended that live attenuated schizont vaccines developed from locally isolated T. annulata strains be used to control the disease. In addition, every care should be taken to prevent introduction of the disease into new areas.

Ticks infesting animals in the Sudan and Southern Sudan: past and current status.

In this review, we collate information about ticks identified in different parts of the Sudan and South Sudan since 1956 in order to identify gaps in tick prevalence and create a map of tick distribution. This will avail basic data for further research on ticks and policies for the control of tick-borne diseases. In this review, we discuss the situation in the Republic of South Sudan as well as Sudan. For this purpose we have divided Sudan into four regions, namely northern Sudan (Northern and River Nile states), central Sudan (Khartoum, Gazera, White Nile, Blue Nile and Sennar states), western Sudan (North and South Kordofan and North, South and West Darfour states) and eastern Sudan (Red Sea, Kassala and Gadarif states).

Comparison of the molecular structure of the TaSP gene of Theileria annulata from Sudanese isolates.

The polymorphic region of the Theileria annulata surface protein (TaSP) was cloned and sequenced from different isolates of cattle and cell lines from different areas of Sudan. Amino acid sequence alignment revealed a high diversity showing amino acid and length polymorphism, both within and between parasite isolates. The generation of TaSP diversity may allow the evasion of host immunity by the parasite since TaSP is a highly antigenic parasite protein.

Epidemiology of Theileria annulata infection of dairy cattle in the Sudan using molecular techniques.

This study provides the first epidemiological data regarding T. annulata infection of diary cattle in Sudan using a combination of routine microscopic examination and two molecular techniques, PCR and reverse line blot (RLB).

Validation of the indirect TaSP enzyme-linked immunosorbent assay for diagnosis of Theileria annulata infection in cattle.

An ELISA based on a recombinant Theileria annulata surface protein (TaSP) was evaluated for detection of antibodies in sera from cattle exposed to tropical theileriosis in Sudan. The reference positive samples, used in this study, were from Theileria-infected populations and consisted of 80 cattle from an endemic area in Khartoum State, with high antibody titers in the indirect fluorescent antibody test (IFAT). The reference negative samples were taken from non-exposed populations and consisted of 120 cattle maintained under strict tick control at a commercial farm in Sudan. The cut-off value determined by Two-Graph Receiver-Operating Characteristic (TG-ROC) curves was set at 31.6%, based on the positive reference samples. Further diagnostic validation was performed, which consisted of the measurement of the area under the ROC (AUC) and by valid range proportion (VRP), which was 0.97 and 0.98 for the cut-off, respectively. There were no cross-reactions with antibodies raised against Babesia spp. It is concluded that the TaSP ELISA is a useful test for the diagnosis of T. annulata infection in cattle under field conditions.