1. Introduction

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The Human Immunodeficiency Virus (HIV), a lentivirus and causal agent of Acquired Immunodeficiency Syndrome (AIDS), was first described in the early 1980s [1]. Based on their gene sequence homology, morphology and life cycle, Lentiviruses are capable of causing latent infection of cells in the long term, or cytopathic effects in the short term, slowly inducing progressive and fatal diseases [2]. There are two types of HIV; HIV-1 and HIV-2 (also Referred to as Lymphadenopathy-Associated Virus type 2), which produce similar clinical syndromes but differ in gene structure and antigenicity. HIV-1 is the most common cause of AIDS in the United States as well as in Brazil, while HIV-2 infection is predominantly found in West African countries [2].

Early HIV infection is the stage of undetected viral load or any clinical manifestation. It is the stage of rapid viral replication, intense immune response and immune destruction. It is also the stage where atypical micro biota has been observed in the gut of an infected individual [3].For example in their investigation they observed overrepresentation of pathogenic Candida albicans and Pseudomonas aeruginosa in early HIV patients.

Those species have been implicated in opportunistic infection observed in HIV infected individuals. In another study, species such as bifidobacteria and lactobacilli, with propensities to boost gut immune function by modulating the innate immune system in pathogenic infection, were observed to be depleted in the faecal microbiota of untreated HIV patients [4].

A shift in proportions of pathogenic and commensal microbes relative to viral load was also observed in another study carried out by Rossit et al, [5] involving early stage HIV patients, either treated or untreated. In this study imbalances in adherent gut bacterial communities leads to significant enrichment and depletion of the Enterobacteriaceae family and Bacteroides class, respectively, in untreated HIV positive subjects compared to treated subjects which portend development of diarrhea. However ART treated HIV patients were observed to exhibited a bacteria composition similar to that of negative individuals.

Gut bacteria normally associated with HIV patients includes the following E. coli, Salminella sp, Shigella sp, Campylobacter jejuni. For example several strains of E. coli have been reported to be associated with persistent diarrhoea in HIV patient. Among the Shigella genus, S. sonnei is of great significance for instance S. sonnei was detected in 11% of diarrhoeic HIV-infected children from the Northwestern region of São Paulo State, Brazil [5]. Approximately the same percentage of Shigella sp. was reported for adult HIV seropositive diarrhoeic individuals from Southern India [6]. Salmonella sp. especially non-typhoidal typies considered another important pathogen for the HIV-infected group as these enterobacteria are 20 to 100 times more frequently isolated among HIV individuals compared to the immunocompetent population [7,8]. Enteric salmonellosis was also more frequently seen in patients in the advanced stages of HIV infection and with impaired nutritional status [9].

2. Materials And Methods

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2.1. Collection Of Sample
A total of 200 stool samples were used for this investigation. One hundred and fifty stool samples were collected from the sero-positive patients attending ART clinic at University Teaching Hospital Ado Ekiti, while fifty stool and blood samples were collected from uninfected individuals (control). The blood samples (4ml) from Suspected HIV negative individuals were collected through finger pricking with sterile lancet and used to screen them for their HIV status. The stool samples from both the HIV positive individuals and the control were subjected to both macrobiological and microbiological investigations. All samples were processed within 24 hours of collection.

2.2. Stool Sample Analysis
The collected stool samples were analysed both macroscopically (colour and texture of the stool e.g., whether watery, form or semi-form) and by culturing. Each stool sample was stirred together before a portion of each was transferred into a new sterile bottle containing peptone water and then homogenized. A loopful of each was then cultured on MacConkey agar except in the case of anaerobic culture where blood agar was used. Selenite F was then added to the remaining sample, incubated at 370C for 24 hours and then subculture on Deoxyckolate Citrate Agar (DCA). For anaerobic culturing Gas jar and pak was used and the samples were incubated for 24 hours at 37oC.

2.3.Isolation Of Bacteria
Both anaerobically and aerobically incubated plates were observed for visible colonies and then sub-cultured on fresh media by streaking method to obtain a pure isolates. The pure isolates were morphologically characterized and subculture into nutrient agar slant for identification and characterization [10].

2.4. Identification And Characterization Of Isolates
The bacteria isolates were identified and characterized based on their colonial morphology, microscopic appearance, grams reactions and biochemical reactions, including carbohydrates utilization profile. The isolates were identified by comparing characteristics with those of known taxa as described by [10].

3. Results

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3.1. Distribution Of HIV Infection Among Patients Used For The Investigation Based On Gender Age
Out of total number of 150 HIV patients attending the ART clinic at EKSUTH, Ado-Ekiti that were recruited for this investigation, 112(74.7%) were found to be females while 38(25.3%) were found to be males. Out of the females sampled, the infection was observed to be higher among the age group 36-45years while it was lowest among the age group 66-75 years. For the male counterpart, the infection was highest in the age group, 26-35 years while it was lowest in the age group < 25 years (Table 1). In general, age group 26-35yerars had the highest incidence of HIV infection among the participants.

Table1

3.2. Types Of Bacterial Species Isolated From The Stool Of HIV Patient And The HIV Negative Individuals Investigated
The following bacterial species were isolated from the stool of the HIV patients used in this investigation. These are Serratia marcescens, Escherichia coli, Salmonella typhi, Citrobacter freundii, Shigella species, Salmonella paratyphi A, Klebsiella pneumoniae, Enterobacter species, Proteus vulgaris, Yersinia enterocolitica, other Salmonella sp., Proteus mirabilis, Providencia species, Pseudomonas aeruginosa, Morganella morganii, Aeromonas species, Staphylococcus aureus, Staphylococcus epidemidis, Streptococci species, Lactobacillus sp and Enterococus sp. Almost the same types of bacterial species were also found in the stool of the control subjects except that Pseudomonas aeruginosa, Morganella morganii, Aeromonas species, Lactobacillus sp and Enterococcus sp were only found in the stool of the HIV patients but not in the stool of the control subjects while Proteus vulgaris was found only in the stool of the control subjects and not HIV patients. All the isolated bacteria had higher frequency of occurrence in HIV patients than the control subject except for Proteus vulgaris that was not found at all in their stool and other Salmonellae sp. that was found only 3(30%) of the HIV patients unlike 7(70%) occurrence found in the stool of HIV negative individual (Table 2).

Table2

3.3. Frequency Of Occurrence Of Bacteria Isolated From Control, HIV Patients On HAART, HIV Patients Not On HAART And The Control
The following bacterial isolates were common in the stool of both HIV patients on HAART,HIV patients who are on HAART and the control individuals investigated. These are Escherichia coli, Salmonella typhi, Shigella species, Enterobacter species, Salmonellae sp, Providencia species, Coagulase positive Staphylococcus and streptococcus sp. While Serratia marcescens, Yersinia enterocolitica, Proteus mirabilis, coaguolase negative Staphylococcu, were only found in the stool of HIV patient on HAART and the control. Also Pseudomonas aeruginosa, Morganella morganii and Lactobacilli sp were only isolated in the stool of HIV patients who are on HAART while Aeromonas species was only found in HIV patients who are not on HAART. (Table3).

Table3

4. Discussion

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In this study, the high prevalence of HIV infection among the female gender observed was in agreement with the report of Mabayoje and Akinleye [11] in the study they carried out among the people in 15 local government areas within Osun State in South West Nigeria. The high prevalence of HIV infection among this gender could be due to the fact that some women may be unaware of their male partner’s HIV status and the gender violence against young females which is on the increase [12]. Also the risk of having HIV infection in female is higher than that of the male because of the anatomic structure of their sexual organ.

Age group 26-35years had highest prevalence of HIV infection (35.3%). This contradict 53% reported in a similar study conducted in West Virginia among age group 35-44years in 2013 Surveillance [13], though higher than 4.4% reported by NARHS Plus II, [14] among age group 35-39 years. This present result however show correlation with recent HIV Surveillance report by United States where prevalence was high among the age group 20-24 years followed by 25-29years [15]. The difference in the result may be due to different geographical location and the age when the individual started having sex. The high prevalence of HIV infection recoded among this age group in this study could be attributed to multiple sex partners and lack of the knowledge of the predisposing factors to this infection.

Escherichia coli (35.5%) accounted for the most frequently encountered bacterial sp. isolated from HIV patients and the HIV negative individuals (control) followed by Klebsiella pneumoniae (10.7%) agree with the report of Marbou, [16]; Fredrick et al. [17]. It also agree with the reported of Samie et al. [18] who worked on the diarrhoeagenic bacterial pathogens in HIV-positive patients in Rural Communities of Limpopo Province, South Africa. The low occurrence of Aeromonas species (0.4%) in this study correlates the result of Hayath et al. [19]who recorded low number of Aeromonas among hospitalized HIV infected patients in southern India.

Moreover, the high occurrence of Pseudomonas aeruginosa 8(100%), Shigella sp (85%), Klebsiella species (83.3%), Salmonella typhi (78.9%), E. coli (78.2%), Salmonella paratyphi A (75%), Serratia marcescens (72.7%), Staphy aureus (66.7%) and Enterobacter Species (57.1%) among HIV positive patients on HAART in this study agrees with report of the work of Yu-Jie et al. [20] who reported that these bacteria were highly enriched in the mucosa of HIV patients, and they are known as opportunistic pathogens and sources of bacteremia in HIV-infected subjects. It also agrees with the report of Estes et al. [21] who observed that there was a persistence of an HIV-associated microbiota in some individuals on long-term ART. The presence of these bacteria among the group could indicate the inability of HAART to total by restore the gut microbiota though this work did not put in to consideration the duration of the antiretroviral therapy of the patients. This observation however is at variance with the report of Obi and Bessong, [22] who recorded a low percentage of these organisms. However, in 2007, Zajac et al. [23] reported that these bacteria shared more than 90% sequences of HIV-1, thus involved in the pathogenesis of HIV.

However, there was limitation to this present study as it was greatly difficult to get equal number of HIV positive individuals who are on HAART and those not on HAART for proper comparison.

5. Conclusions

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In this study it was observed that HIV infection was significantly higher among the females than males and among traders than other occupations investigated. It was also observed to more prevalent in the age group 26-35 years.

The types of bacteria isolated from the stool of |HIV positive individuals on HAART were almost similar to those found in HIV negative individuals. However there was a higher percentage of most of the pathogenic bacteria such as E. coli, Shigella species, Salmonella typhi, Klebsiella pneumoniae, Salmonella paratyphi A, Yersinia enterocolitica, Proteus mirabilis, Enterococcus spp, Streptococcus sp, and Providencia species among HIV patient. In addition to the ones that were found only in HIV patient screened which are Pseudomonas aeruginosa, Morganella morganii, Aeromonas species and Enterococcus spp.

Recommendation

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1. Awareness of HIV infection in relation to causative agent, mode of transmission and the risk factor is highly imperative in the community sampled in order to reduce the incidence rate of the infection

2. Combination of chemotherapy and nutrition is advocated to manage HIV infection.

3. Antibiotic should be prescribed only after pathological investigation of types of pathogenic microorganisms being harboured by HIV positive individuals to prevent indiscriminate use of antibiotic.

4. Early administration of HAART to HIV positive patients is highly recommended. This will help preventing the rapid replication of the virus.

5. A research into probiotic therapy is recommended to that can help reconstitute the shift in the gut microbiota

6. Contribution To Knowledge

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This study has shown that although HAART is very important, it is not enough for the management of HIV infection, because the administration of HAART did not have significant effect on bacterial population in the stool of HIV positive individuals as compared to the bacterial population in the stool of control individuals with lower population, which is a reflection of the immune status of these seropostive individuals.

This research work however has shown that the bacteria isolated from HIV seropositive individuals are more sensitive to antibiotics than bacteria isolated from the HIV negative individuals.

Moreover, the investigation showed higher prevalence of the infection in the female gender than the males It also showed that the highest prevalence of the infection was found in age group 26-35 years and it is also prevalent among traders.

References

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