Kudi A.A., Saminu C.M., Hassan M.D., Manga M.M:

Department of Medical Microbiology & Immunology Federal Teaching Hospital, Gombe
Deborah B., Madina E.K.,

Centre for Integrated Health Programme (CIHP), Gombe Regional Office.
Sa’idu A.

Department of Medicine, Federal Teaching Hospital, Gombe

All correspondence to: kudiayuba@gmail.com

ABSTRACT
A success story at the Federal Teaching Hospital, Gombe (FTHG) a 500-bed capacity tertiary institution situated at the heart of the North-eastern Nigeria. A manually operated PCR facility was first installed by a supporting partner – the Institute of Human Virology-Nigeria (IHVN) in 2009. The FTHG in collaboration with Centre for Integrated Health Programme (CIHP) and Centre for Disease Control (CDC) – Nigeria up-graded the facility to an Automated PCR machine (COBAS TAQMANN96) in 2015. It was a good response to partnership that demonstrates programme commitment and sustainability. In that spirit, human and material resources were committed towards the actualization of space and infrastructure with quality finishing/furnishing of the PCR Laboratory. Staff were highly motivated and the dearth in Laboratory infrastructure was greatly addressed. Timely Supply of reagents and consumables was also sustained. The resultant benefit of this partnership was quality improvement to services with increased in patients’ access to HIV and Hepatitis B and C diagnosis and treatment monitoring. Our Centre remains ever grateful to the PEPFAR HIV programme, our supporting partners and the Federal Ministry of Health, Nigeria.

Key Words: PCR, PEPFAR HIV and AIDS Programme, Partnership.

INTRODUCTION

Gombe, a State located in the Northeast region of Nigeria within the expansive savannah. It shares border with Yobe, Taraba, Bauchi, Borno and Adamawa States. The State has an area of about 20, 265 square kilometres and a population of around 2,353,000 people as at 2006 census.1 Gombe has 2 distinct climatic conditions, the dry season (November to March) and the raining season (April to October) with an average rain fall of 850mm.1

The Federal Teaching Hospital Gombe (FTHG) is located in Gombe, a tertiary 500-bed capacity health institution situated in the heart of the North-eastern Nigeria, relatively equipped with states of the arts facilities in Diagnostic Laboratory, addresses wide range of investigations. Before the coming of the Centre for International Health Programme (CIHP) as a supporting partner in the field of HIV investigations and treatment monitoring, the Laboratory could only boast of a manually operated PCR equipment installed by the Institute of Human Virology, Nigeria (IHVN)] in 2009, under the Presidential Emergency Plan for AIDS Relief (PEPFAR), a programme of the United States of America launched in 2003 to combat
global HIV/AIDS.2,3 For us at the Federal Teaching Hospital Gombe (FTHG), Nigeria, the up-grading of the PCR Laboratory to automation by the CIHP/CDC (Centre for Disease Control) – Nigeria in partnership with FTHG management was a success story. The Hospital Management, staff and people of Gombe State will ever be grateful to PEPFAR.

Team work and Positive Response to Partnership
The News to install an automated PCR Machine (COBAS TAQMANN96) came to us (Laboratory Management) as an institution, and was observed that our supporting partner, CIHP were serious, as ever; they placed the card on the table that; PCR automated equipment could be installed in our facility to boost the Centre’s capacity in HIV and AIDS investigations. What was required then of the FTHG Management to put in place were space (infrastructure) and few other logistics. The laboratory Team saw the possibility because of the close working relationship, support and co-operation being received at all times from the Hospital management.

In high Spirit, the Laboratory leadership Team led by the Director Laboratory Services, approached the Hospital management with a proposal to that effect, and sooner than expected, discussion with the management for theconstruction of a brand new PCR Laboratory was concluded and approved. The Chief Medical Director (CMD), Dr. Abubakar Sa’idu assured the leadership of CIHP that construction of the PCR Laboratory, furnishing and any other necessary item(s) needed according to specification to enable the centre have the automated PCR equipment installed will be put in place. He equally made a strong commitment towards the deadline given by CIHP/NCDC- Nigeria for the space to be ready. To his words, within 3 months (April to July, 2015), all were set for PCR equipment installation. On 1st October, 2015, PCR equipment with all the necessary accessories for Early Infant Diagnosis (EID) and Viral quantification (viral load) reagents arrived the FTHG. PCR Equipment (COBAS TAQMANN96) and all the accompanied accessories were received by the Director Laboratory services in good deadline given by CIHP/NCDC- Nigeria for the space to be ready. To his words, within 3 months (April to July, 2015), all were set for PCR equipment installation. On 1st October, 2015, PCR equipment with all the necessary accessories for Early Infant Diagnosis (EID) and Viral quantification (viral load) reagents arrived the FTHG. PCR Equipment (COBAS TAQMANN96) and all the accompanied accessories were received by the Director Laboratory services in good condition and according to the specifications earlier given.

On 13th October, 2015, installation and on-site training of staff were completed by a Team of Roche Engineers with the support of the CIHP Gombe Laboratory Team, led by the Senior Laboratory Supervisor, Mrs. Deborah Bulus. The arrival of -20oC and -80oC deep freezers from the CIHP supporting partners, and the installation of 20Kva inverter by same (CIHP) later, climaxed the infrastructural up-grading of the PCR Laboratory at the FTHG Centre. The now fully equipped PCR Laboratory was commissioned by the Hon. Minister of Health, Prof. Isaac F. Adewole on 11th January 2016. This has greatly addressed the dearth in Laboratory infrastructure, improves the quality of services in the Centre and increased access; staff were also highly motivated to quality issues.

To further galvanized its commitment to quality and demonstrate a kind of strong partnership, aside from EID and HIV (viral load) investigations fully supported by the CIHP/CDC-Nigeria, the FTHG management added Hepatitis B and Hepatitis C viral quantification to the menu of investigations using the installed PCR machine. So far, from the date the PCR machine was installed to the time of writing this report, more than 36,840 samples were received from Kaduna, Benue and Gombe States for HIV viral load, 5,157 for EID, while 242 for Hepatitis B virus
and 85 for Hepatitis C virus were analysed. The addition of these tests to the HIV investigations have greatly enhanced quality of patient care and reduced the hardship hitherto been experienced by the patients.
CONCLUSION
The FTHG Management and staff have not only demonstrated good response and sensitivity to partnership, but also understood the principle and importance of programme sustainability. This was clearly seen in the resources being committed, both human and material towards the actualization of space and infrastructure with quality finishing/furnishing of the PCR Laboratory. This is our success story. We remain ever grateful to our programme partners and the Federal Ministry of Health. We look forward to more of supports and such types of collaboration from well-meaning groups and organisations in and outside Nigeria.

REFERENCES

1. Gombe State action plan of His Excellency, 29th May, 2011 to 29th May, 2015. An official document of the Gombe State Government.

2. Partnering to Achieve Epidemic Control in Nigeria. www.pepfar.gov

3. PEPFAR HIV and AIDS programme on the Nigeria

Ojih M.A
Department, of Medical Microbiology Ahmadu Bello University Teaching Hospital Zaria.
Amadi Y.H
Department of Parasitology and Entomology Ahmadu Bello University Zaria.
Akpulu S.P
Department of Human Anatomy Ahmadu Bello University Zaria.
Yaro E.G
Department of Human Anatomy, Ahmadu Bello University, Zaria, Nigeria

All correspondence to: E-mail: maryarichayakubu@gmail.com

ABSTRACT
Background: Intestinal parasites and helminthic infection has posed medical changes to the developing word. There is therefore need to source and develop local drugs or herbs to combat this menace. Aims and Objectives: This study therefore examined the anthelminthic activity of ethanolic leaf extract of combretum molle in vitro on the ova of Hookworm and Haemonchus contortus. Materials and Methods: One drop of the extract mixed 5 drops of distilled water was added to 6 drops of faecal samples on a microtiter well and incubated for 14 days at 260C, using water as control. The activity was assessed by comparing the number of ova present after a period of 2 weeks for both the extract and water. The result showed that there was only one egg (non -embryonated) of hookworm observed in the well with the extract. For the Haemonchus contortus in the well with extract, there as one egg,12 unembryonated eggs, 13 dead larvae. For the control(water), there was no positive effect on the hookworm and the Haemonchus contortus. Conclusion: Result from this study shows that ethanolic leaf extract of combretum molle was effective against the ova of the hookworm and the Haemonchus contortus and can be a possible anthelmentic agent.

Keywords: helminthic, Haemonchus contortus. combretum molle, Hookorm

Introduction

Intestinal parasites and helminthic infection has posed medical changes to the developing word. Resistance to antibiotics is becoming a serious problem worldwide. The discovery of new and effective antimicrobials and/or resistance modulators is necessary to tackle the activity of spread of helminths. Infections with helminths are among the most widespread diseases worldwide and the reason for considerable morbidity, mainly in populations from less developed countries of Sub-Saharan Africa, the Americas, and Southeast Asia (Hotez et al.,2008). Recent investigations estimated the number of people infected with the most common parasitic nematodes Ascaris lumbricoides, Trichuris trichiura, and hookworms (Ancylostoma duodenale and Necator americanus) to be more than 1.7 billion worldwide, including 100 million infections in West Africa (Pullan et al., 2010). Extensive measures have been undertaken by the WHO to provide effective and low-cost treatments for the inhabitants in endemic areas, which have successively led to a decrease in the morbidity and prevalence of intestinal helminthiases (Humphries et al.,2012). In most cases, albendazole is the deworming drug of choice, however, mebendazole, pyrantel, and levamisole are also classified as essential medicines (WHO,2013). Nevertheless, the number of approved anthelmintic drugs the societies are relying on is very limited and especially large-scale preventive applications bear the risk of resistances to emerge (Vercruysse et al.,2011). In veterinary medicine, this is already the case where worldwide resistances of parasites of livestock have increased to a dramatic degree within the last years (Kaplan and Vidyashankar,2012). Despite the availability of conventional drugs, traditional medicine, often based on medicinal plants, plays a crucial role in the treatment of helminthic infections. As indicated by a recent field study, many African patients prefer to consult traditional healers or herbal practitioners for this indication (Agyare et al.,2009). More than 300 species of plants have been reportedly used in traditional control of human and animal helminthosis in Africa (Nwude et al.,1998). Combretum molle of the combretacae family is used to treat human ailments including abdominal discomfort, body pains, respiratory discomfort, cods and fevers, ear and eye ailments, schistosomiasis, hookworms, dysmenorrhea and infertility in women, leprosy, syphilis, microbial infections and general body weakness (Ademola and Eloff, 2010). Helminthiasis is a major limitation to the livestock industry in Africa. Haemonchus contortus is the singular most important helminth responsible for major economic losses in small ruminants. The high cost of anthelmintics to small farmers, resistance to available anthelmintics and residue problems in meat and milk consumed by humans further complicates matters.

Haemonchus contortus is the single most important constraint to sheep production in South Africa (Vatta et al.,2001) with estimated direct and indirect economic losses of US $45 million (I.G. Horak personal communication quoted by Waller, (2006). The cost of controlling helminth infestation is also very high globally with chemotherapy remaining the most widely used method of treatment. Unfortunately, the excessive use of these drugs when not necessary, in addition to their use at incorrect doses, has resulted in the wide scale emergence of resistance in this parasite. The use of plants and plant extracts as a possible source of new anthelmintic has received more interest in the last decade. This study therefore aim to study the anthelminthic activity of Combretum molle (velvet bush willow) against ova of hookworm and Haemonchus contortus.

Materials and methods
This study was conducted in the department of parasitology and entomology, faculty of veterinary medicine Ahmad Bello University, Zaria. Positive stool sample was collected from Ahmad Bello University Teaching Hosital Zaria. Plant collection. Five-kilogram leaves of Combretum molle was collected in Zaria and identified in the biological science department of Ahmad Bello University, plant extraction. The leaves were air dried (290C) under shade and grinded sing pestle and mortar into fine powder as described by Onyeyili et al.,2001. 150g of the powdered sample was extracted with 2.5L of 96% ethanol using cold maceration (percolation) method. The filtrate was evaporated and the extract obtained used. One part of Acacia was mixed with 5 parts of the extract.one drop of the mixture as introduced into microtiter well for culture and left for two weeks using water as control. 0.1ml distilled water was added to 1.2ml positive faecal sample and 0.2ml extract. Culture was observed for ova microscopically on day 7, 10 and 14 post incubation respectively.

Discussion
The results from this study shows that at the end of 14 days, the extract at 0.2ml had an adverse effect on the ova of Hookworm and the egg, and larvae of Haemonchus contortuus. This was observed by the absence and dead of the ova and larva. which implies that the extract has anthelminthic effect. While in day 7 and 10, the extract did not reduce the number of the ova and larvae significantly the control only with water did not show any anthelminthic effect rather, the ova hatched to larvae.

These findings are similar with previous investigations on lethal anthelmintic effects against a variety of nematode species, including adult C. elegans Mohamed et al (2000) and L3 larvae of Toxocara canis Kiuchi et al (2000). Also, the activity against C. elegans (L4 larvae/ young adults) of a plant extract from Combretum mucronatum by Schum. & Thonn. (Combretaceae) leaves has been confirmed in vitro (adult worms) and in vivo against Trichuris muris and against adult stages of A. ceylanicum, Spiegler et al., (2015) while the same extract was inactive against third stage larvae of the hookworm Kone et al., (2012). This inactivity is probably caused by the sheath protecting the free-living L3 larva persevering in soil, which is unlikely to be penetrated by large polar molecules such as proanthocyanidins. T. et al., (2012). Similarly, Adamu et al (2013) reported that the plants extract generally had better inhibition activity on the eggs than on the larva of H. contortus based on the EC50 values recorded. This is contrary to report by Ademola where they had better activity for the larval inhibition compared to the egg hatch assay.

In comparison to work undertaken using extracts of other plant species, the activity shown by the three plants with the best activity were in the same order of activities found by Bizimenyera et al., (2006) using Peltophorum africanum leaf, bark and roots as well as extracts of Coriandum sativum on eggs and larval development of H. contortus Eguale et al.,(2007). Bizimenyera et al., (2006) recorded an EC50 of 0.62 mg/ml and 0.72 mg/ml for the leaf acetone extract for the EHA and LDT assays respectively. This is in the same order with values of 0.62 and 0.64 mg/ml recorded in the EHA and LDT for H. trifoliata in the current study. The EC50 of H. trifoliata was lower than the acetone leaf extracts of Combretum molle 0.87 mg/ml and 0.60 mg/ml for the EHA and LDT respectively (Eloff McGaw and Eloff, 2008). The weak activity recorded by M. azedarach (EC50 = 10.96 mg/ml) agrees with Maciel et al., (2006) in Brazil where they reported an LC50 of 9.18 mg/ml with the leaf ethanol extract of the plant. It is encouraging that despite the difference in geographical location and organic extractants used, similar bioactivity was obtained for related species.

Conclusion
This finding shows that Combretum Molle has an Anthelminthic activity on the Ova of Hookworm and Haemonchus 

Acknowledgment
We are grateful to the Management and Laboratory staff Ahmadu Bello University Teaching Hospital Zaria, and Department of Parasitology and Entomology, Faculty of Veterinary Medicine Ahmadu Bello University for permission, support and cooperation during the work.

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