Chapter 1: Introduction

Tiger nut milk, locally known as kunun aya, is a traditional non-alcoholic beverage

popular across Nigeria, particularly in the northern region, where it has long been

valued for its sweet flavor and health benefits. Derived from Cyperus esculentus,

commonly known as tiger nut, this drink has been consumed for centuries and

continues to be a preferred choice among locals for its purported nutritional and

medicinal properties. It is typically made by blending soaked tiger nuts with water,

followed by straining to obtain a creamy liquid that serves as a natural, lactose-free

alternative to cow’s milk.

Within the context of the Niger State Polytechnic Zungeru campus, kunun aya is

widely consumed by both students and staff. The beverage is often sold by local

vendors due to its affordability, refreshment value, and perceived health benefits.

However, as with many locally processed foods, there are increasing concerns

regarding the microbiological safety of the milk due to poor hygiene during

processing and inadequate storage conditions.

Microbial contamination can occur at various stages, from harvesting the tiger nuts

to preparing and selling the milk. If these contaminants, such as Escherichia coli,

Staphylococcus aureus, and fungi like Aspergillus species, are not properly

controlled, they pose significant health risks to consumers. Furthermore, the

physiochemical properties of the milk, such as its pH, moisture content, and fat

content, influence both its nutritional quality and shelf life. A lack of proper

physiochemical assessment can lead to the consumption of low-quality or even

harmful milk.

This study aims to assess the microbiological and physiochemical quality of locally

processed tiger nut milk sold within the Niger State Polytechnic Zungeru campus.

By examining both its microbial load and physiochemical characteristics, the study

seeks to identify potential health risks and provide recommendations for improving

the safety and quality of kunun aya.

Research Questions:

1. What are the common microbial contaminants present in locally processed

tiger nut milk?

2. Are the physiochemical properties of kunun aya within acceptable ranges for

safe consumption?
 3. What measures can be implemented to reduce contamination and improve

the quality of tiger nut milk?

Aims and Objectives:

i. To determine the microbiological safety of tiger nut milk by identifying and

quantifying bacterial and fungal contaminants.

ii. To evaluate the physiochemical properties, such as pH, moisture content, fat

content, and protein content, to assess the milk’s nutritional value and shelf

life.

iii. To suggest possible interventions and improvements in processing and

storage practices for vendors.

Scope of Study: This study will focus on the vendors selling kunun aya within the

Niger State Polytechnic Zungeru campus. A variety of samples will be collected

from different vendors, ensuring that a broad representation of the milk quality is

analyzed. The study will be limited to microbiological and physiochemical

assessments, with recommendations aimed at improving the local processing

practices of tiger nut milk.

Significance of Study: The results of this study will provide critical insights into the

safety and quality of tiger nut milk, which has become a popular beverage within

the campus community. By identifying microbial hazards and highlighting the

physiochemical attributes of the milk, this research will contribute to improving

public health standards in the area. Additionally, the study’s findings may be of

interest to food safety regulatory bodies such as NAFDAC, helping to shape policies

for ensuring safer food and beverage processing methods in Nigeria.

Chapter 2: Literature Review

2.1 Tiger Nut (Cyperus esculentus)

Tiger nuts, though classified as tubers, are often mistaken for nuts due to their size and

appearance. The plant, Cyperus esculentus, belongs to the sedge family and is commonly

cultivated in regions of Africa, particularly Nigeria, where it grows in both wild and

cultivated settings. It is valued for its edible tubers, which have a sweet, nutty flavor, and

can be consumed raw, roasted, or processed into milk.

Historically, tiger nuts have been a staple food in many cultures due to their resilience in

harsh climates and their dense nutritional profile. In Nigeria, they are particularly noted

for their high fiber content, which aids digestion, and their abundance of vitamins and

minerals, including vitamin E, magnesium, and potassium. Tiger nuts also contain
beneficial fats, predominantly monounsaturated fatty acids, which contribute to

cardiovascular health.

2.2 Processing of Tiger Nut Milk

The traditional process of making tiger nut milk involves soaking dried tiger nuts in water

to rehydrate them. Afterward, the nuts are blended with water to extract the liquid. This

liquid is then strained to remove any solid particles, yielding a creamy, dairy-free milk.

Variations of the recipe may include flavoring with sweeteners like dates or spices such

as ginger, cinnamon, or cloves, depending on local preferences.

In many parts of Nigeria, including Niger State, this process is carried out at a small,

local scale by vendors who often lack the necessary infrastructure for stringent hygiene

control. This exposes the milk to possible contamination, especially when vendors use

untreated water, unclean utensils, or sell the milk in open-air environments. Given the

high moisture content and nutrient-rich nature of the milk, it is highly susceptible to

microbial growth, making proper storage and handling practices essential for safety.

2.3 Microbial Contamination in Foods

Microbial contamination is a major concern in food processing, particularly in

environments with limited sanitation. In the case of kunun aya, several studies have

identified various bacteria, including Escherichia coli, Salmonella, and Staphylococcus

aureus, as potential contaminants. These pathogens can cause foodborne illnesses ranging

from mild gastrointestinal discomfort to severe, life-threatening conditions.

Contamination can occur at multiple points in the production process. Poor hygiene

during harvesting, storage of tiger nuts, inadequate washing of nuts, or the use of

contaminated water can introduce harmful microorganisms. Additionally, the ambient

temperature in Nigeria, which is conducive to bacterial growth, further exacerbates the

problem if the milk is not refrigerated or consumed within a short period after

preparation.

2.4 Physiochemical Properties of Beverages

Physiochemical properties such as pH, moisture content, total solids, ash content, and

nutrient composition are crucial indicators of a beverage’s quality and stability. In the

case of tiger nut milk, the pH tends to be slightly acidic (typically between 4.5 and 6.0),

which helps to preserve its freshness but can also promote the growth of acid-tolerant

bacteria if not properly handled.

Moisture content is another key factor influencing the shelf life of the milk. Higher

moisture content increases the risk of microbial growth, which is why proper drying and

storage of the raw tiger nuts are essential. The fat and protein content of kunun aya also

contribute to its nutritional value, offering a source of essential fatty acids and plant-

based proteins.
Several studies on the physiochemical properties of plant-based milk alternatives have

highlighted the variability in these properties depending on the processing methods used.

For instance, processing temperatures, blending times, and filtration methods can

significantly alter the nutrient profile and shelf life of the milk. Therefore, assessing these

parameters in locally processed tiger nut milk is critical for understanding its quality.

2.5 Food Safety Practices

Globally, food safety regulations emphasize the importance of hygienic practices in food

preparation to prevent contamination and ensure the safety of consumers. In Nigeria,

regulatory bodies such as the National Agency for Food and Drug Administration and

Control (NAFDAC) provide guidelines on food safety standards. However, many local

vendors, especially those in informal sectors like campus markets, often operate outside

the purview of these regulatory bodies, increasing the risk of contamination.

The implementation of good manufacturing practices (GMP) and hazard analysis and

critical control points (HACCP) systems can mitigate these risks by identifying critical

points in the production process where contamination is most likely to occur and

instituting controls to prevent it. While such systems are commonplace in industrial food

processing, they are rarely employed by small-scale vendors.

Chapter 3: Methodology

3.1 Study Area

The study was carried out within the Niger State Polytechnic Zungeru campus, located in

Zungeru town, Niger State, Nigeria. The campus is home to a diverse student population

and hosts a variety of vendors selling food and beverages, including kunun aya. The

vendors were selected based on their accessibility and the popularity of their products

among students and staff.

3.2 Sample Collection

3.2.1 Sampling Locations Samples of tiger nut milk were collected from five different

vendors within the Niger State Polytechnic Zungeru campus. These vendors were

selected to ensure diversity in processing methods, storage conditions, and potential

contamination sources.

3.2.2 Sample Size A total of 25 samples were collected, with five samples taken from

each vendor at different times of the day (morning, afternoon, and evening) to account for

potential variations in contamination levels due to temperature fluctuations and handling

practices.

3.2.3 Collection Method Sterile glass containers were used to collect the tiger nut milk

samples from each vendor. These containers were properly sealed and labeled with the

vendor’s identification, collection time, and date. The samples were immediately
transported to the microbiology laboratory under cold storage conditions (4°C) to prevent

further microbial growth during transport.

3.3 Microbiological Analysis

The microbiological analysis aimed to detect and quantify common bacterial and fungal

contaminants that could compromise the safety of the tiger nut milk. The following tests

were performed on each sample:

3.3.1 Total Viable Count (TVC) The total viable count was conducted to determine the

overall bacterial load in each sample. This was performed using the plate count method:

Procedure: Serial dilutions of the milk samples were prepared in sterile distilled water.

From each dilution, 0.1 mL was plated on nutrient agar and incubated at 37°C for 24 to

48 hours. The colonies that formed were counted, and the total viable count was

calculated as colony-forming units per milliliter (CFU/mL).

3.3.2 Coliform Count Coliform bacteria are indicators of fecal contamination and poor

hygiene during food handling. The presence of coliforms in tiger nut milk was assessed

using MacConkey agar:

Procedure: 0.1 mL of each milk sample was plated on MacConkey agar and incubated at

37°C for 24 hours. The appearance of pink colonies indicated the presence of lactose-

fermenting coliform bacteria. The coliform count was expressed in CFU/mL.

3.3.3 Pathogen Identification To identify specific pathogens, selective media were used:
Escherichia coli: Eosin Methylene Blue (EMB) agar was used to detect E. coli. Colonies

with a metallic sheen were indicative of E. coli.

Salmonella species: Xylose Lysine Deoxycholate (XLD) agar was employed to isolate

Salmonella species. The presence of red colonies with black centers was recorded as

Salmonella positive.

3.3.4 Fungal Count To assess the presence of fungal contaminants, Potato Dextrose Agar

(PDA) was used:

Procedure: 0.1 mL of each sample was plated on PDA and incubated at room temperature

(25°C) for 3 to 5 days. Colonies of yeast and mold were identified and counted.

3.4 Physiochemical Analysis

The physiochemical properties of the tiger nut milk were evaluated to determine its

quality and nutritional value. The following parameters were assessed:

3.4.1 pH Measurement The pH of the milk samples was measured to determine their

acidity or alkalinity. The pH can influence the taste, microbial growth, and shelf life of

the milk:

Procedure: A calibrated pH meter was used to measure the pH of each sample. The pH

values were recorded to the nearest 0.01 unit.

3.4.2 Moisture Content The moisture content indicates the amount of water present in the

milk, which affects its texture, shelf life, and microbial growth:
Procedure: Approximately 10 mL of each milk sample was dried in an oven at 105°C

until a constant weight was achieved. The moisture content was calculated as the

percentage loss in weight relative to the original sample weight.

3.4.3 Total Solids The total solids content represents the combined amount of all solid

components in the milk, such as fats, proteins, and carbohydrates. It is an important

indicator of the milk’s nutritional value:

Procedure: After determining the moisture content, the remaining dried sample was

weighed to calculate the total solids percentage.

3.4.4 Ash Content The ash content is a measure of the mineral content of the milk:

Procedure: The dried samples were incinerated in a muffle furnace at 550°C for 2 hours.

The remaining ash was weighed and expressed as a percentage of the total sample

weight.

3.4.5 Fat Content The fat content of the milk was measured using the Soxhlet extraction

method:

Procedure: Approximately 5 mL of each milk sample was subjected to Soxhlet extraction

using hexane as the solvent. The fat content was determined by the weight of the

extracted fat relative to the sample size.

3.4.6 Protein Content The protein content was assessed using the Kjeldahl method, which

involves the digestion of the sample, followed by distillation and titration:

Procedure: Each sample was digested in sulfuric acid with a catalyst. The digested

solution was distilled, and the released ammonia was titrated with a standard acid

solution. The protein content was calculated based on the nitrogen content of the milk.

3.5 Data Analysis

The data obtained from the microbiological and physiochemical analyses were subjected

to statistical analysis to identify trends and significant differences between samples:

Statistical Software: The data were analyzed using SPSS (Statistical Package for the

Social Sciences) version 25.0.

Analysis: Descriptive statistics, including means, standard deviations, and frequency

distributions, were calculated. Inferential statistics such as ANOVA (Analysis of

Variance) were used to compare results across different vendors and sample times.

3.5.1 Ethical Considerations

All vendors were informed of the purpose of the study and gave verbal consent for

sample collection. The study was conducted following the ethical guidelines for food

safety and microbiological research. Personal and vendor identities were anonymized in

the study to maintain confidentiality.

Chapter 4: Results and Discussion

4.1 Microbiological Results

The microbiological analysis was conducted to assess the microbial load and presence of

specific pathogens in tiger nut milk. The results are summarized in Table 4.1 and

discussed below.

Test Range of Results (CFU/mL)

Total Viable Count (TVC) 10^4 - 10^6

Coliform Count 10^2 - 10^4

Escherichia coli Isolation Detected in 2 samples

Salmonella Isolation Not detected

Fungal Count 10^3 - 10^5

4.1.1 Total Viable Count (TVC)

The total viable count, which measures the overall bacterial load in the milk samples,

ranged from to CFU/mL. According to the food safety guidelines for non-pasteurized

beverages, a bacterial load of CFU/mL or higher is considered potentially hazardous for

human consumption. The high bacterial load in these samples suggests that many of the

vendors may be processing and handling the tiger nut milk under substandard hygienic

conditions.
4.1.2 Coliform Count

Coliform bacteria were detected in 18 out of the 25 samples, with counts ranging from to

CFU/mL. The presence of coliform bacteria is a strong indicator of fecal contamination,

most likely due to poor handling and hygiene practices. These results are concerning, as

coliform bacteria can cause gastrointestinal infections and other health issues in

consumers.

4.1.3 Pathogen Identification

Out of the 25 samples tested, Escherichia coli was detected in two samples. E. coli is a

pathogenic bacterium that indicates fecal contamination and poses a significant health

risk to consumers, potentially causing foodborne illnesses. No Salmonella species were

detected in any of the samples, which is a positive outcome, as Salmonella is a common

cause of food poisoning.

4.1.4 Fungal Count

The fungal count ranged from to CFU/mL. The presence of yeasts and molds in the

samples suggests that spoilage organisms are prevalent, which may result from improper

storage conditions and prolonged exposure to environmental contaminants. Although

yeasts and molds are not necessarily pathogenic, they can degrade the quality of the milk

and cause it to spoil quickly.

4.2 Physiochemical Results

The physiochemical analysis was conducted to evaluate the quality and nutritional value

of the tiger nut milk. The results are summarized in Table 4.2 and discussed below.

4.2.1 pH

The pH values of the tiger nut milk samples ranged from 4.5 to 5.5, indicating slight

acidity. The pH of a beverage can influence its stability and microbial growth. Tiger nut

milk with a pH below 4.6 is considered less susceptible to bacterial growth, but the

results suggest that most samples fall within a range that may support the growth of

spoilage organisms, particularly at higher storage temperatures.

4.2.2 Moisture Content

The moisture content of the milk samples ranged from 82% to 88%, which is consistent

with the high water content of tiger nut milk. This makes the beverage susceptible to

rapid spoilage, especially if it is not stored under proper refrigeration conditions. The

high moisture content provides a suitable environment for microbial growth, which aligns

with the microbiological findings of high bacterial and fungal counts.

4.2.3 Total Solids

The total solids content, which includes fat, protein, carbohydrates, and other nutrients,

ranged from 12% to 18%. This variation could be due to differences in the processing

methods used by different vendors, particularly the amount of water added during

preparation. The total solids content is a key factor in determining the nutritional value of

the milk.

4.2.4 Ash Content

The ash content, which represents the mineral content of the tiger nut milk, ranged from

0.5% to 1.0%. This suggests that the beverage contains a moderate amount of essential

minerals, such as calcium, potassium, and magnesium. Minerals contribute to the overall

nutritional benefits of tiger nut milk.

4.2.5 Fat Content

The fat content of the samples ranged from 3% to 5%, which is consistent with the

natural fat content of tiger nuts. Fat is a critical source of energy and contributes to the

creamy texture of the milk. Tiger nut milk is a nutritious alternative to cow’s milk,

especially for those who are lactose intolerant.

4.2.6 Protein Content

The protein content of the samples ranged from 2% to 4%. Proteins are essential for the

growth and repair of body tissues. Although tiger nut milk is not as high in protein as

some other plant-based milks, it still provides a valuable source of nutrition.

4.3 Discussion

The microbiological analysis revealed that locally processed tiger nut milk sold within

the Niger State Polytechnic Zungeru campus is prone to microbial contamination,

particularly with coliform bacteria and Escherichia coli. These findings raise concerns

about the safety of the beverage, as fecal contamination and poor hygiene during

processing are likely contributors to the high microbial load. The presence of yeasts and

molds also indicates the potential for spoilage, particularly under inadequate storage

conditions.

The physiochemical analysis showed that tiger nut milk is rich in moisture, fat, and

minerals, making it a nutritious beverage. However, the high moisture content and

slightly acidic pH create favorable conditions for microbial growth, which can

compromise the shelf life and safety of the milk if not properly handled.

Overall, the results suggest that while tiger nut milk offers significant nutritional benefits,

there are considerable risks associated with its microbiological safety. Implementing

stricter hygiene practices during processing and ensuring proper storage conditions are

essential to minimizing microbial contamination and improving the overall quality of the

milk.
Chapter 5: Conclusion and Recommendations

5.1 Conclusion

The microbiological and physiochemical assessment of locally processed tiger nut milk

(kunun aya) sold within the Niger State Polytechnic Zungeru campus has provided

valuable insights into the safety and quality of this popular beverage. The findings

revealed significant microbial contamination, particularly with coliform bacteria and

Escherichia coli, suggesting poor hygiene practices during processing and handling.

Additionally, the high fungal counts indicated a risk of spoilage, especially under

inadequate storage conditions.

From a nutritional perspective, tiger nut milk was found to be rich in essential nutrients,

including fats, proteins, and minerals. However, the high moisture content and slightly

acidic pH present challenges in maintaining its quality over time. Without proper

refrigeration and hygienic handling, the milk is prone to spoilage and microbial growth,

which could pose health risks to consumers.

In conclusion, while tiger nut milk offers numerous health benefits, the safety concerns

identified in this study must be addressed to ensure its safe consumption.

5.2 Recommendations

Based on the findings of this study, the following recommendations are made:

1. Improvement of Hygiene Practices: Vendors should be educated on proper

hygiene practices during the processing, handling, and packaging of tiger nut milk

to reduce the risk of microbial contamination.

2. Regular Microbiological Testing: Regular microbiological testing should be

conducted to monitor the safety of tiger nut milk sold within the campus and

ensure compliance with food safety standards.

3. Proper Storage Conditions: Vendors should be encouraged to store tiger nut milk

under refrigerated conditions (below 4°C) to minimize microbial growth and

extend its shelf life.

4. Public Health Awareness: Public health campaigns should be conducted within

the campus to raise awareness among consumers about the potential health risks

of consuming improperly processed tiger nut milk.

5. Further Research: Additional research should be conducted to explore the effects

of different processing techniques on the microbiological safety and nutritional

value of tiger nut milk.

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