Shawgi M. Hassan

Entomogenous fungi as promising biopesticides for tick control.

When ticks were sealed in nylon tetrapacks and infected with the entomogenous fungi, Beauveria bassiana and Metarizium anisopliae and maintained in potted grass in the field, the fungal oil formulations (109 conidia per ml) induced 100% mortality in larvae of Rhipicephalus appendiculatus and Amblyomma variegatum, whereas mortalities in nymphs varied between 80–100% and in adults 80–90%. The aqueous formulations (109 conidia per ml) induced mortalities of 40–50% and reductions in egg hatchability of 68% (B. bassiana) and 48% (M. anisopliae) when sprayed on Boophilus decoloratus engorging on cattle. The strains of B. bassiana and M. anisopliae isolated from naturally infected ticks were also found to induce high mortalities in both R. appendiculatus and A.variegatum in tetrapacks placed in potted grass. Both aqueous and oil-based formulations were found to be effective, although the latter induced higher mortalities. These fungal strains in aqueous formulation (108 conidia per ml) suppressed on-host populations of adult R. appendiculatus by 80% (B. bassiana) and 92% (M. anisopliae) when sprayed on tick-infested grass once per month for a period of 6 months. The feasibility of using entomogenous fungi for tick control in the field is discussed.

The impact of Ixodiphagus hookeri, a tick parasitoid, on Ambl yomma variegatum (Acari: Ixodidae) in a field trial in Kenya

In an experiment to investigate the effect of the tick parasitoid, Ixodiphagus hookeri, on tick numbers on cattle, 150 000 parasitoids were released over a period of 1 year in a field where ten cattle infested with multiple tick species were kept. Amblyomma variegatum was reduced from 44 to two ticks per animal while Rhipicephalus appendiculatus increased over the time of parasitoid release. During the time of release 51% of the nymphs of A. variegatum collected from the animals were parasitized. The recovery of the parasitoids after the releases were stopped was only 9%. The total numbers of A. variegatum remained low up to 1 year after the parasitoid release was stopped. This study gives an insight into how I. hookeri could be used strategically for the management of A. variegatum on small-scale farms.

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.

Diagnosis of Babesia caballi and Theileria equi infections in horses in Sudan using ELISA and PCR

The purpose of this study was to estimate the prevalence of equine piroplasmosis in Sudan. The presence of antibodies against Babesia caballi and Theileria equi was determined in serum samples obtained from 158 horses raised in different locations in Sudan by enzyme-linked immunosorbent assay (ELISA). The B. caballi 48-kDa and the T. equi EMA-2 purified recombinant proteins were used as antigens in the ELISA test. Results showed that seven (4.4%) were positive for B. caballi and 80 (63.5%) were positive for T. equi. Polymerase chain reaction (PCR) assays have been applied using primers targeting the B. caballi 48-kDa merozoite antigen, the T. equi SSUrRNA and the T. equi EMA-1 genes. PCR performed on 131 blood spots in filter paper revealed that 33 (25.2%) samples were positive for T. equi but no positives were found for B. caballi. It is concluded that equine piroplasmosis is endemic in the country. This is the first study on serological and molecular epidemiological diagnosis on equine piroplasmosis in Sudan.

Efficacy of Rhipicephalus (Boophilus) microplus Bm86 against Hyalomma dromedarii and Amblyomma cajennense tick infestations in camels and cattle

The recombinant Bm86-based tick vaccines have shown their efficacy for the control of cattle ticks, Rhipicephalus (Boophilus) microplus and R. annulatus infestations. However, cattle ticks often co-exist with multi-host ticks such as Hyalomma and Amblyomma species, thus requiring the control of multiple tick infestations for cattle and other hosts. Vaccination trials using a R. microplus recombinant Bm86-based vaccine were conducted in cattle and camels against Hyalomma dromedarii and in cattle against Amblyomma cajennense immature and adult ticks. The results showed an 89% reduction in the number of H. dromedarii nymphs engorging on vaccinated cattle, and a further 32% reduction in the weight of the surviving adult ticks. In vaccinated camels, a reduction of 27% and 31% of tick engorgement and egg mass weight, respectively was shown, while egg hatching was reduced by 39%. However, cattle vaccination with Bm86 did not have an effect on A. cajennense tick infestations. These results showed that Bm86 vaccines are effective against R. microplus and other tick species but improved vaccines containing new antigens are required to control multiple tick infestations.

Epidemiology of Theileria annulata Infection of Dairy Cattle in the Sudan Using Molecular Techniques

Abstract:  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).

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.

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.

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.

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).