Asian Journal of Healthy and Science

p-ISSN: 2980-4302

e-ISSN: 2980-4310

Vol. 2 No. 1 January 2023

 

 

MICROBIOLOGICAL QUALITY OF HERBAL FORMULATION USED FOR THE TREATMENT OF TYPHOID FEVER SOLD IN MAKURDI METROPOLIS, CENTRAL NIGERIA

 

Aernan P.T.1, Odo J.I.2, Sar T.T.3, Nweke O.A.4

Department of Microbiology, Federal University of Agriculture P M B 2373, Makurdi, Benue State, Nigeria1,3,4

Department of Fisheries and Aquaculture, Federal University of Agriculture P M B 2373, Makurdi, Benue State, Nigeria2

Email: odojoel@gmail.com

Abstract

Typhoid, dysentery and cholera. To investigate the microbiological quality of herbal formulations. Herbal formulations were purchased from four different markets (Wadata market, Wurukum market, Modern market, and North bank market) in Makurdi metropolis Microbiological analysis was carried out using pour plate isolation method. Identification of isolated microorganisms was based on their cultural, morphological, biochemical characteristics using standard microbiological procedures. Microbiological analyses showed that the total bacterial counts (TBC) of all the test herbal samples obtained from the various markets ranged from 1.8 x103 to 9.3x103 cfu/ml and the total fungal count in the herbal mixture had a range of 1.0x103 to 2.5x103, Four bacterial species were identified and they include; Bacillus spp, Escherichia coli, Staphylococcus aureus and Enterobacter spp. The least occurring bacterial isolates was Bacillus spp (12.5%), while the highest occurring was Staphylococcus aureus (37.5%). Four fungal isolates were identified and they include, Aspergillus niger, Penicillium spp, Scedosporium spp and Phialophora parasiticum. Aspergillus niger and Phialophora parasiticum were the least occurring fungal isolate (12.5%) while Scedosporium spp and Phialophora parasiticum were the most occurring fungal isolate (37.5%). Since applications of herbal medicines for curative purposes is on the increase, there is a need for risk assessment of microbial load of the medicinal plants at critical control points during processing. Furthermore, the danger associated with the potential toxicity of herbal therapies employed over a long period of time demand that the practitioners be kept abreast of the reported incidence of renal and hepatic toxicity resulting from the ingestion of medicinal herbs.

Keywords: Herbal Formulation; Typhoid Fever; Bacterial Counts; Bacillus Spp; Toxicity.

 

 

 

INTRODUCTION

It is estimated that approximately 80% of the population in developing countries uses herbal medicine as part of their primary health care. The use of herbs dates back to the time of the early man, who had crudest tools as his implements and used stones to start his fire (Kafani, 1994). The medicinal science came into existence through a Greek man called Hippocrates and as such he earned his reputation as father of medicine (Heber, 2008). Most savage people have believed that disease was due to the presence of evil spirits in the body and could be driven out only by the use of poisonous and disagreeable substances calculated to make the body an unpleasant place to remain. The knowledge regarding source and use of the various products suitable for the purpose was usually restricted to medicine men of the tribe (Sharma, 2006)

Contamination of herbal medicine product is defined as the “undesired introduction of impurities of a chemical or microbial nature or of foreign matter, into or onto a starting material, intermediate product or finished product or finished flavored product throughout production, sampling, packaging, or repackaging, storage or transport”

   The introduction of micro-organisms in the herbal medicinal products can alter the physicochemical characteristics of the products, which may lead to harmful effects to the quality of the herbal medicine products. The organisms he said were Staphylococcus aureus, Eschericha coli, Rhyzopus stolonifer, Candida species, etc (Adeleye et al., 2005).Bacteria are ubiquitous in nature, therefore because of the ubiquitous nature of bacteria, they contaminate almost every material, herbal juice no exception (Ochei.,2007).

Among the main safety risks related to medicine is contamination by micro-organisms of various kinds that may be adherent in leaves, stems, flowers, seeds, and roots from which herbal medicines are prepared. Alternatively, microorganisms can be introduced during harvesting, handling, open air drying, preserving, and manufacturing.

Although the medicinal plants with their chemical constituents carry huge applications in the treatment or prevention of various diseases. The plant materials are exposed to various contaminants like toxic elements, pesticides, residues etc. but the chief contaminants mainly responsible for the deterioration of herbal products are the microbe. Several studies showed that herbal plants are associated with broad variety of microbial contaminants (kneifel et al., 2002). When herbal products contaminated with these micro-organisms are consumed by people, they can cause serious health issues, one of the most serious issues surrounding the safety of herbal formulations concerns regulating the level of mycotoxins found in herbal preparations. Mycotoxins are a group around 400 toxic secondary metabolites produced by fungi such as Aspergillus,  penicillium, Fusarium, Claviceps and Alternaria  Recently Nyarko et al.(2007) showed that 22% of herbal plant samples studied failed to comply with the quality requirement for traditional medicine.

Herbal medicine is most often polyherbal, being prepared from mixtures of different plant parts obtained from various plant species and families and may contain multiple bioactive constituents that could be difficult to characterize (Ogbonnia et al., 2010). The bioactive principles in most herbal preparations are not always known and there could be possibilities of interaction with each other in solution. The quality as well as the safety criteria for herbal drugs may be based, therefore, on a clear scientific definition of the raw materials used for such preparations. Also herbal medicine may have multiple physiological activities and could be used in the treatment of a variety of disease conditions. (Yidana and Bayorbor, 2002 ) It could be administered in most disease states over a long period of time without proper dosage monitoring and consideration of toxic effects that might result from such prolonged usage (Ogbonnia et al., 2010).

The usage of herbs as medicines is determined mostly by the community and environment in which one grows up. Addo (2007) carried out a study to determine the socio-demographic characteristics and pattern of use of herbal medicines by women admitted to the Obstetrics and Gynaecology Department in the Komfo Anokye Teaching Hospital (KATH), a teaching hospital serving the Northern part of Ghana and made the following observations: More than fifty percent (50%) of patients used herbal medicines which were mostly unknown to the attending health workers. The less educated as well as the unskilled/ semi-skilled used herbal medicines more frequently compared to their more skilled and educated counterparts. Herbal medicine use is thus more prevalent in the groups who usually have poor socio-economic facilities and carry most of the burden of social deprivation. It is possible that their disease conditions may be adversely affected.

To achieve the desired benefit from herbal preparations, an individual must take the required dose over a certain length of time. Although it is generally believed that most herbal preparations are safe for consumption, some herbs like most biologically active substances could be toxic with undesirable side effects (Bisset, 1994).

 

RESEARCH METHODS

The study was carried out in Makurdi, Benue State. It has a population of 226,198 (1991 census) with a landmass of 16km radius. It has major market like North Bank market, Wurukum market, High level market, Modern market and Wadata market.

Commercially sold herbal formulation was purchased from four different markets (North bank market, Wurukum market, Modern market, and Wadata market). Two samples were purchased from each market and were immediately transported to the Department of Microbiology laboratory of the Federal University of Agriculture, Makurdi for microbiological analysis.

 

RESULT AND DISCUSSION

Table 1 Presents the Morphological and bacterial counts of each sample in 4 different markets in Makurdi metropolis. It shows that MM1 (Modern market 1) had the highest counts of 12.5x103 cfu/ml, while WD2 (Wadata 2) had the least count of 1.7x103.

 

 

 

 

 

 

 

Table 1. Morphological and colony count of bacteria isolated from herbal formulation from 4 different markets in Makurdi metropolis

Sample source                                     Markets                                                 Counts (cfu/ml)

 

Herbal formulation                                  NB1                                                       9.3X103

 

Herbal formulation                                  NB2                                                       3.4X103

 

Herbal formulation                                  WD1                                                      4.2X103

 

Herbal formulation                                  WD2                                                      1.7X103

 

Herbal formulation                                  WK1                                                      2.5X103

 

Herbal formulation                                  WK2                                                      1.8X103

 

Herbal formulation                                  MM1                                                      12.5X103

 

Herbal formulation                                  MM2                                                      1.9X103

 

Table 2 Shows the average counts of fungal isolates which shows that NB1 (North bank 1) had the highest count of 2.5x103 cfu/ml, while the least count was obtained from WD1 (Wadata 1) with 1.0x103 cfu/ml.

 

 

 

 

 

 

 

 

 

 

 

 

 

Table 2. Average colony count of fungal isolates from herbal formulation in 4 different markets in makurdi metropolis

Sample source                                       Market                                         colony count(cfu/ml)

 

Herbal formulation                                  NB1                                                       2.5 X103

 

Herbal formulation                                  NB2                                                       2.0 X103

 

Herbal formulation                                  WD1                                                      1.0 X103

 

Herbal formulation                                  WD2                                                      2.0 X103

 

Herbal formulation                                  WK1                                                      2.3 X103

 

Herbal formulation                                  WK2                                                      1.4 X103

 

Herbal formulation                                  MM1                                                      1.5 X103

 

Herbal formulation                                  MM2                                                      1.8 X103

 

Table 3 and 4 shows the occurrence of bacterial isolates from the sample collected from four different market. Staphylococcus aureus had the highest frequency of 37.5% of the isolates in the sample. Bacillus spp had the lowest frequency of 12.5% of the bacterial isolates in the sample. (North Bank 1) and WK2 (Wurukum 2) had all the bacterial isolates recorded in this study from herbal formulation, while WD1 (Wadata 1)  had only 1 of bacterial isolate.

 

 

 

 

 

 

 

 

Table 3. Occurrence of bacterial isolates from herbal formulation in 4 different markets in makurdi metropolis

Isolates                                                   Frequency                                           Percentage

Staphylococcus aureus                            3                                                           37.5%

 

Entermoeba spp                                       2                                                           25%

 

Escherichia coli                                       2                                                            25%

 

Bacillus spp                                             1                                                            12.5%

 

Table 4. Occurrence of the bacterial isolates from the different markets

Sample         Markets          Staphylococcus    Enterobacter     Escherichia         Bacillus    

Source                                   aureus                    spp                      coli                        spp

 

Herbal           NB1                    +                            +                       +                             +

Formulation  

                       NB2                    +                            +                       -                               -

                       

                       WD1                   +                            -                         -                              -

                          

                       WD2                   -                            +                        +                              +        

 

                       WK1                    +                            -                       -                               +

 

                       WK2                   +                             +                      +                              +

 

                       MM1                  +                             -                        +                              +

 

                       MM2                   +                             -                        +                              +

                    

-          = Absent

+     = Present

 

The percentage occurrence of fungal isolates from the herbal formulation, shows that Scedoporium spp and Penicillium spp both had the same frequencies of occurrence representing 37.5% of the total fungal isolates. While Aspergillus spp, had a percentage frequency 12.5% of the total fungal identified. (Table 5)

 

 

 

Table 5. Percentage occurrence of fungal isolates from herbal formulation in 4 different markets in Makurdi metropolis

Isolates                                               Occurrence                                                Percentage

Scedosporium spp                                3                                                                  37.5%

 

Penicillium  spp                                    3                                                                  37.5%

 

Aspergillus niger                                  1                                                                  12.5% 

 

Phialophora parasiticum                       1                                                                 12.5%

 

On table 6 the occurrence in percentage of fungal isolates in four different markets is presented. The study shows that WK2 (Wururkum 2) had all the fungal isolates in the herbal formulation identified representing the highest frequency of occurence, while MM1, (Modern market 1) had had the least fungal isolates.

 

Table 6. Occurrence of fungal isolates from herbal formulation in 4 different markets in Makurdi metropolis

Sample             Market             Scedosporium           Penicillium       Aspergillus        Phialophora

Source                                      spp                           spp                   spp                    spp

Herbal              NB1                 +                               -                      -                                 +

Formulation      

                          NB2                 -                                +                      +                                +

 

                          WD1                -                                -                       -                                 +

 

                          WD2                +                               +                       +                                 -

 

                          WK1                +                               -                       +                                 +

 

                          WK2                +                               +                       +                                +  

 

                          MM1                -                                +                        -                                -

 

                          MM2                +                              +                        +                                -

-           = Absent

+         = Present

 

This study has shown that there are varieties of microorganism present in our various herbal formulation which could have resulted from contaminated soils, plants and its products, preparation processes, quality of water, containers and processing equipment. However, these microorganisms exhibit multi-resistance to many antibiotics. Since herbal formulations are mainly prepared for human consumption, there is a very high chance of passing the antibiotics resistant microorganisms into the human ecosystem. This poses a great danger to human health. Since herbal concoctions are prepared using varieties of medicinal plants which contain active constituents that are cheap and effective against common bacterial infections.

A number of sources of contamination of herbal preparation especially during preparation have been identified. The microflora of the final product may represent contaminants from the raw materials, equipment, water, and atmosphere and from personnel. Microorganisms such as Escherichia coli, and Scedosporium  spp reported in this study are generally known to proliferate in potable, distilled and de-ionized water while Bacillus spp, Staphylococcus, Aspergillus and penicillin are commonly isolated from air. The most common source of post preparation herbal contamination is the packaging vessels. 

In order to enhance consumer acceptability, most herbalists in Nigeria have adopted the use of bottles and plastic containers as packaging vessels for their preparations. Unfortunately these vessels are not subjected to any form of sterilization after washing them. Contamination of the preparation coupled with the humid tropical environments may result in the proliferation of microbial contaminants in the herbal remedies (WHO, 2007); this probably explains the high microbial counts recorded in this study. Such high levels of microbial contamination have been shown to result in spoilage and degradation of the products or may constitute a health hazard to the user. (Akande et al., 2013) Most herbal preparations are made up of different components of various plant species and these preparations are not standardized with respect to color, taste, consistency etc.

Unlike the orthodox drugs, changes in the appearance, odor, taste etc of herbal formulations due to spoilage are hardly readily detected by the patients. (Acharya et al., 2008) Among the microorganisms isolated from the Herbal Medicine, Bacillus, Staphylococcus, E. coli and penicillin were the major contaminants. Although the pathogenicity of these organisms was not assessed, species of these agents have been incriminated in serious human infections. Bacillus spp are widely distributed in the soil, dust, air and water and they are resistant to environment destructive factors. Apart from the unacceptable microbial loads observed in the samples, the presence of contaminants considered to be completely unacceptable in herbal preparations was demonstrated. The most common isolates in the tested samples were Gram-positive organisms belonging to the genera Bacillus and Staphylococcus aureus are normal commensals of the mammalian skin, hands and mucous membranes.

Upon the consideration of the extent of human contact involved in the preparation of herbal medicinal samples, it is most likely that sources of the contaminating Staphylococcus spp. are the producers of the herbal formulations this suggests that the level of hygiene of persons involved in the preparation of the tested samples may be low. Similar studies carried out on herbal samples include work by Akande et al. (2013); Alakali et al.  (2016); Oluyege et al.(2010); Okeniyi et al. (2005); Oreagba et al. (2011); Adeleye et al. (2005) have all reported that the pathogens frequently isolated in herbal products were S. aureus, E. coli. Except for Scedosporium which was not isolated in some reports mentioned above. This work varies by reporting a higher count of Bacillus spp. Contamination by Bacillus spp could have arisen during growth of the herbs as the bacterium is commonly found in soils. Escherichia coli, a major faecal coliform may have been introduced from the water used during processing of the herbs. Microbial contamination of  herbal mixture as shown in this study, may also be as a result of the plant materials, utensils used during preparation, poor hygiene of the manufacturer or even the packaging vessel after processing as also suggested by many authors earlier repoted.

Microorganisms are present everywhere and can easily contaminate any substrate. Considering the packaging materials, it is worthy of note that this contributes greatly to microbial contamination as the final stage of the processing is packaging. Most of the packaging cans used are not sterilized and are usually picked up where they are found littered along the road or in public places and barely washed before being used to package finished herbal products. It was observed that fungal growth was more than bacterial growth and this is attributable to the low pH value of all the herbal formulation samples which is favorable for fungal growth. The high microbial load and presence of specific pathogens in the tested herbal formulation heave serious clinical as well as pharmaceutical implications. Clinically, consumers of any of these products are at risk of contracting infections by the different pathogens which may be of great consequence if not identified and treated appropriately. Presence of Eschericha coli in the sample indicates poor hygiene practices and lack of adequate handling of the products.

According to the European pharmacopoeia 2007, no Salmonella spp or Eschericha coli strain should be present in samples (Oluyege and Adelabu, 2010). The intake of a high concentration of accumulated toxins produced by organisms such as Bacillus cereus, Staphylococcus aureus and Aspergillus niger may lead to undesirable reactions in consumers. Olusegun (2007) reported that the presence of Bacillus spp. in herbal preparations is an indication that the water used in the preparation of the products is not from a good source. The risk is greater if the consumer is a young child with undeveloped immunity, an elderly with diminished immunity or the immunocompromised patients. Incidentally, these groups of consumers are the most in need of herbal medicines for the treatment of many diseases to which they are susceptible.

 

CONCLUSION

Staphyococcus aureus, Escherichia coli, Enterobacter spp, Bacillus spp Scedosporidum spp Penicillium spp, Aspergillus niger, and Phialophora parasiticum spp   were isolated and identified as micro- organisms associated with contamination of herbal formulations sold in markets within Makurdi metropolis, with Staphylococcus aureus having the highest bacterial colony count and Scedosporium spp having the highest fungal count.

 

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Copyright holders:

Aernan P.T., Odo J.I ., Sar T.T, Nweke O.A (2023)

First publication right:

AJHS - Asian Journal of Healthy and Science

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