QUALITY INDICATORS OF SOME FRESH WATER FISH MARKETED IN SHARKIA PROVINCE, EGYPT

CONCLUSION:

The present study concluded that the examined fish species were contaminated bacteriologically by aerobic microorganisms and family enterobacteriaceae with varying counts, and chemically by histamine which affect its bacteriological, chemical quality and nutritive value. Therefore, strict hygienic measures should be considered in order to achieve good quality fish.

QUALITY INDICATORS OF SOME FRESH WATER FISH MARKETED IN SHARKIA

PROVINCE, EGYPT

AlaaEldinM. Morshdy ;Abdallah F. Abdallah

andEsraa A. Abdelaziz

Food Control Department, Faculty of Veterinary Medicine,

Zagazig University, Zagazig 44519, Egypt.

Key Words: Bacteriologicalquality,Clariaslazera, Histamine, Oreochromisniloticus.

ABSTRACT

The presented study was conducted to evaluate the organoleptic, bacteriological and chemical quality ofsome fresh water fish species (Oreochromisniloticusand Clariaslazera) marketed in Zagazig city, Sharkia governorate, Egypt. The results revealed that,the organoleptic evaluation of O. niloticusand C. lazera sampleswas 94.2% and 90.7%.Themean values of total bacterial count in the examinedOreochromisniloticusandClariaslazerasamples were 6.63±0.06 and 6.58±0.057log₁₀ CFU/g,while, themean values of total enterobacteriaceaecount were 4.5±0.048 and 4.8±0.084 log₁₀ CFU/g,respectively. Histamine was found in 45(90%) of O. niloticus but all C. lazerasamples contained histamine with mean values of 10.7±2.5 and 16.01±3.24mg ̸100g inO. niloticusand C. lazera.

INTRODUCTION

Fish is a highly perishable, very rich protein diet that involves high levels of free amino acidsas well as volatile nitrogen bases essential for human consumption in addition to minerals, vitaminsand polyunsaturated fatty acid (Korkmaz et al., 2019). Oreochromisniloticus (Tilapia) is fresh water, fast growing fish farmed mainly in Africa and Asia. Tilapia fish are essential for human because they constitute a major part of human diet and provide our bodies with the needed proteins(Ghorbani and Mirakabad, 2010). Clariaslazera (catfish) have been of great interest to many producers because of their efficient feed conversion and fast growth rates (Ali and Jauncey, 2005).

Bacteriological quality of fish is very important to the public health because it is responsible for fish spoilage and subsequent food poisoning due to complex processes of bacterial degradation which result in deterioration of fish. Also, these bacteria have the ability to metabolize the amino acids producing ammonia and biogenic amines including putrescine, histamine, cadaverine, organic acids, ketones and sulfur compounds. The Aerobic plate count (APC) is as an indicator of bacterial populations in fish. Enterobacteriaceae is a widely distributed family in nature and gastrointestinal tract of fish which is related to human infection; microorganisms of this family are Gram-negatives (Kaper et al., 2004).

Foodborne microorganisms can cause many disease outbreaks, causing health risks to consumers (Rohde et al., 2014). The greatest risk to human health occurs due to the consumption of raw, inadequately cooked or insufficiently processed fish, and fish products. The symptoms of fish food poisoning include nausea, vomiting, diarrhea, abdominal cramps, headache as well as muscle and joint pain. Contamination of fish with organisms of public health significant remains primarily a problem of handling and processing (WHO, 1999).

Quality of fish is often more difficult to control due to variations in species, sex, age, habitats and action of autolytic enzymes as well hydrolytic enzymes of microorganisms on the fish muscle (Venugopal, 2002). Therefore, surveillance of potential contaminants in fish is very critical to ensure safety of consumers. For fish industry, freshness assurance is considered as one of the most important goals because freshness is closely related to quality. Fish quality is of a major concern to the food processors, consumers, and public health. Provision of safe, wholesome and acceptable fish and fish products, control of contamination is very essential.

Histamine is a heterocyclic alkaline biogenic amine formed in fish by microbial decarboxylation of free amino acids. Scombroidpoisoning results from ingestion of fish contains high levels of histamine and other amines which potentiate histamine (Ayesh et al., 2012). Scombroid fish poisoning is an acute illness, symptoms include sweating rash, facial flushing, abdominal cramps and diarrhea but more severe symptoms include swelling of the tongue and throat, respiratory distress and blurred vision which require medical treatment (Hwanget al.,2011). Therefore,the present study was planned to throw a light on theorganoleptic, bacteriological and chemical quality of some fresh water fish (Oreochromisniloticus and Clariaslazera)sold at Zagazig markets, Sharkia governorate, Egypt.

MATERIALS AND METHODS

1. Collection of samples:

A total of one hundred samples of raw fish (Oreochromisniloticusand Clariaslazera, 50, each) were randomly collected from different fish mrketsatZagazig city, Sharkia Governorate. The samples were aseptically transferred in an ice box to Meat Hygiene Laboratory, Faculty of Veterinary Medicine, Zagazig University for organoleptic, bacteriological and chemical examination.

2. Organoleptic examination:

Organoleptic examinations of fish were carried out to detect any abnormal changes in different parts of fish body and its physical characters. This examination wascarried out according to the procedure recommended by Braumuller (1958) according to Table (1).

Table 1: Organoleptic examinations of fish.

The result was presented as follow 50% or more:  fit for human consumption; 25%-50%: border line (rapid consumption); and less than 25%: decomposed (unfit).

3. Bacteriological examination

3.1. Preparation of samples:

Fish samples based on its side over sterile plate hold by sterile forceps, surface sterilized by hot spatula and removed by sterile forceps.  Samples will be prepared according to the technique recommended by ICMSF (1978) as following: - Under complete aseptic condition, 10 grams of fish sample will be transferred into sterile homogenizer jar containing 90 ml of sterile peptone water 0.1%. The contents will be homogenized for 2 minutes at room temperature to provide a homogenate of 10^-1 dilution. One ml of homogenate will be transferred into sterile test tube containing 9 ml of 0.1% peptone water, and then ten-fold serial dilutions will be prepared.                                     

3.2. Total bacterial count:(FDA, 1998).

3.3. Total enterobacteriaceae count:     (ICMSF, 1978).

3.3.1. Isolation and identification of enterobacteriaceae:according to Cowan and Steel (1974) and ICMSF (1978).

4. Chemical examination

4.1. Determination of histamine by ELISA:

Quantitative determination of histamine was done by enzyme immunoassay in combination with supplementary kit (cat. no. BA E-1100), according to Leszczynskaet al. (2004).

RESULTS

Organoleptic examination:

The results recorded in Table 2showed that all the examined samples were considered fresh and fit for human consumption from organoleptic standpoint of view. The organoleptic evaluation of O. niloticussampleswas 94.2%, while, it was 90.7% for C. lazera samples.

Table2: Organoleptic examination of fish samples (n=50, each).

n: Number of examined fish samples (50, each).

The examined fish samples were organoleptically fit for human consumption (Total more than 50%).

Bacteriological examination:

Themean values of total bacterial count in the examinedOreochromisniloticusandClariaslazerasamples were 6.63±0.06 and 6.58±0.057log₁₀ CFU/g,with minimum values of 5.54 and 5.61log₁₀ CFU/g, and maximum values of 7.24 and 7.19 log₁₀ CFU/g, respectively. There was non-significant differences (p>0.05) between the two species in total bacterial count (Table3).

As illustrated in Table3, themean values of total enterobacteriaceaecount in the examinedOreochromisniloticusandClariaslazerasamples were 4.5±0.048 and 4.8±0.084 log₁₀ CFU/g,with minimum values of 3.81and 2.70log₁₀ CFU/g, and maximum values of 5.52 and 5.57log₁₀ CFU/g, respectively.

Table3: Statistical analytical results of total bacterial count and Enterobacteriaceae in the examined fish samples (n=50, log₁₀ CFU/g).

n= number of examined samples

CFU / g: colony forming unit per gram

S.E: standard error of mean.

Means of total bacterial count are not significantly different (P > 0.05).

Means of total Enterobacteriaceae (a-b) are significantly different (P ˂ 0.05).

Chemical examination

Histamine was found in 45(90%) of O.niloticus but all C. lazerasamples contained histamine. It ranged from 1 to 22.7 and 1.2 to 31.9 mg ̸100g, with mean values of 10.7±2.5 and 16.01±3.24mg ̸100gin O.niloticusand C.lazera.Non-significance differences between the two species were detected(Table 4).As found in Table (5), 40(80%) and 30(60%) of O.niloticusand C.lazera were accepted.

Table 4:Histamine level in the examined fish samples (n= 50, each).

n: Number of examined fish samples (50, each). S.E: Standard error of mean.

Table 5: Acceptability of the examined fish according to their histamine levels (n=50, each).

n: Number of examined fish samples (50 of each), MRL *: Maximum Residual Limit stipulated by EOS (2010).

Results obtained from Table 6 illustrated the correlation coefficient of relations between the bacteriological and chemical profile of the examined O. niloticus. The correlations between total bacterial count, Enterobacteriaceaecount and histamine in the examined O. niloticussamples were positive weak (0.2 and 0.12), while, the correlations between Enterobacteriaceaecount and histamine were positive weak (0.25).

The correlation coefficient of relations between the bacteriological and chemical profile of the examined Clarias lazera was recorded in Table 6. The correlations between total bacterial count, Enterobacteriaceaecount and histamine in the examined C. lazerasamples were positive weak (0.27and 0.16). However, the correlations between Enterobacteriaceaecount and histamine were negative weak (-0.09) in the examined C. lazerasamples.

Table 6: Correlation between total bacterial count, enterobacteriacae, and histamine level of OreochromisniloticusandClariaslazera(n=50, each).

P. Correlation: Pearson Correlation.

0-0.3: weak correlation, 0.4-0.6: medium correlation, and 0.7-0.9: Strong correlation.

DISCUSSION

Organoleptic evaluation of fish is very important from the consumer point of view. The texture of fish muscles vary from firm to tender. This variation between the samples of O. niloticus and C. lazerareferred to the difference in their catching, handling, storage, transportation, distribution and marketing of methods. Nearly similar results were recorded by Ammar (2001), Morshdy et al. (2002), Abd-Allah (2004) andRaafat (2004),but lower values of organoleptic evaluation were reported by Abo Samara (2001) whofound the average of organoleptic examination of T. nilotica and C. lazera was 80.8% and 83.5%,Aref (2006) who found that the mean values of organoleptic examination of Tilapia spp and C. lazera was 67.5% and 75%.

Total bacterial count has been used to assess the bacteriological load, sanitary quality and safety of fish. There was non-significant differences (p>0.05) between the two species.

Nearly similar result was reported by Mohammed et al. (2017). Lower results of total bacterial count were reported by El-Shabasy (2009) who found themean values of total bacterial count in the examinedO. niloticusand C. lazerasamples were 4.9 and 5.1 log CFU/g, El-Betar (2011) who recorded 4.6 and 6.1log CFU/g as mean values of total bacterial count in the examinedO. niloticusand C. lazera, Prabakaran et al. (2011) who found themean value of total bacterial count was 5.6log CFU/g. However, higher values of total bacterial count (2 to 9.7, 3.9 to 7.9 and 6.8 to 7.9 log CFU/g) were reported by El–Habib (2011), Obemeata et al. (2011) and Mustafa et al. (2013).

The Enterobacteriaceae count is regarded as an important index of fish quality. There was a significant difference (p˂0.05) between OreochromisniloticusandClariaslazerasamples.

Nearly similar result was reported by Mohammed et al. (2017). Lower results were reported by Popelka et al. (2014) who found that Enterobacteriaceae count was 2.1  log CFU/g and Hussein et al. (2019) who foundthemean Enterobacteriaceaecount in the examinedC. lazerasamples was 3.9 log CFU/g.

Moreover, Enterobacteriaceae was determined as a part of the microbial flora during storage of fish under refrigerated storage. The presence of Enterobacteriaceae in fish is due to the widespread of this family in the environment and the natural habitat in the gastrointestinal tract of warm-blooded animals. Enterobacteriaceae may play an important role in the fish spoilage and some bacterial species of this family are very pathogenic (Guiraud, 2003; Shabarinath et al., 2007 and Lopez Da Silva et al., 2010).

The high level of histamine is probably related to bacterial decarboxylase activity due to quality of raw material, miss handling or other causes during their shelf-life (Helmy et al., 2018). So, eating fish with high histamine level leads to acute illness called scombroid fish poisoning which characterized by facial flushing, sweating, rash, diarrhea and abdominal cramps that usually resolve after several hours without medical intervention, but severe symptoms include respiratory distress, swelling of the tongue and blurred vision that need medical treatment (CDC, 2007). Concerning to histamine concentrations, lower results (1-1.5 mg%) were detected in Taiwan by Hwang et al. (2011) and 1.7 mg% by Jaksic et al. (2017). However, Takahashi et al. (2015) reported high histamine content (>50mg%) and 21.59 and 18.31 for C.gariepinusand O.niloticus(Helmy et al. (2018).According to the maximum residual limits (20mg%) proposed by EOS (2010), 80% and 60% of O. niloticus andC. lazerawere accepted.

Histamine is related to scombroid fish poisoning when its concentration exceeds 200 ppm, often above 500 ppm.The disease condition caused by consumption of contaminated fish by performed scombrotoxin is called scombrotoxin poisoning(USFDA, 2001). The main characteristic symptoms of scombrotoxin poisoning are nausea, vomiting, diarrhea, burning in or around the mouth and throat, rashes on the upper part of the body, decrease the blood pressure, headache, dizziness, itching of the skin, respiratory distress, heart palpitation, and. These symptoms occur within a few minutes to a few hours of consumption and last from 12 hours to a few days(EC, 2003).

CONCLUSION:

The present study concluded that the examined fish species were contaminated bacteriologically by aerobic microorganisms and family enterobacteriaceae with varying counts, and chemically by histamine which affect its bacteriological, chemical quality and nutritive value. Therefore, strict hygienic measures should be considered in order to achieve good quality fish.

REFERENCES

Abd-Allah, M. A. (2004): Occurrence of bacterial pollution indicators in Boulti fish (T. nilotica) in Alexandria. Alex. J. Vet. Sci., 22(1): 195-205.

Abo Samara, R. G. (2001): Hygienic evaluation of some fishes marketed in Damietta M. V. Sc. Thesis Department of Food Control (Meat Hygiene), Faculty of vet. Med., Zagazig Univ., Egypt.

Ali, M. Z. and K. Jauncey (2005): Approaches to optimizing dietary protein to energy ratio for African catfish Clariasgariepinus (Burchell, 1822). Aquacul.Nutr., 11: 95–101.

Ammar, M. A. M. (2001): Sanitary assessment of some common fresh water fish in Assiut M. V. Sc. Thesis Fac. Vet. Med., Assiut Univ., Egypt.

Aref, A. M. (2006): parasites of public health importance among marine and fresh water fishes Ph. D. Thesis Fac. Vet. Med., Zagazig Univ., Egypt.

Ayesh, A.;M.Ibraheim; A. El-Hakim and E.Mostafa (2012): Exploring the contamination level by biogenic amines in fish samples collected from markets in Thuel Saudi Arabia. Afr. J. Microbiol. Res., 6: 1158-1164.

Braumuller, H. (1958): Der wert der keimzahlung fur die frischebeurleilung van see fischen Inavg. Diss., Berlin.

Center of Disease Control and Prevention "CDC" (2007):Scombroid fish poisoning associated with tuna steaks. Louisiana and Tennessee, 32: 817- 819.

Cowan, S.T. andK.J. Steel (1974): Manual for the Identification of Med.  Bacteria 2nd Ed. Cambridge Univ., Press, England.

Egyptian Organization for Standardization and Quality Control "EOS" (2010): Maximum Levels for certain contaminants in foodstuffs. No 7136/2010. Egyptian Standards, Ministry of Industry, Egypt.

El-Betar, O. M. (2011):  Quality assessment of some fresh fish. M. V. SC. Thesis, Meat Hygiene, Fac. Vet. Med., Alexandria Univ.

El-Habib, R. (2011): Bacteriological quality and biogenic amines determination by HPLE in Bassa fish imported to Saudi Arabia. Int. J. Pharmacy and Pharmaceutical Scis, 3: 343-347.

El-Shabasy, N. A. (2009): Microbiological Analysis Of Farm Fishes In Alexandria Governorate. Ph. D. V. SC. Thesis,Meat Hygiene, Fac. Vet. Med., Alexandria Univ.

European Commission "EC" (2003): Commission recommendation concerning a coordinated program for the official control of foodstuffs for (2003/10/EC, D J 7I:76-81). BrusselsI.

Food and Drug Administration "FDA" (1998): Environmental chemical contaminants and pesticides. Fish and fishery products hazards and controls guide.

Ghorbani, M. and Mirakabad, H. Z. (2010): Factors influencing on trout production in KhorasanRazavi Province. Trends Agric. Econ., 3: 19-27.

Guiraud, J. P. (2003): Microorganisms intervening in the food industry. Food Ind. J., 1:611-614.

Helmy, N. A.; Hassan, M. A.; Hassanien, F. S. and Maarouf, A. A. (2018): Biogenic amines in seafood. Benha Vet. Med. J., 34(3): 279-286.

Hussein, M.A.; A.M.Merwad ; M.T.Elabbasy ; I.I.Suelam ; A.M.Abdelwahaband M.A.Taha (2019): Prevalence of Enterotoxigenic Staphylococcus aureus and Shiga Toxin Producing Escherichia coli in Fish in Egypt: Quality Parameters and Public Health Hazard. Vector Borne Zoonotic Dis., 19(4): 255-264.

Hwang, C.; H.Kung; C.Lin; D.Hwang and Y.Tsai (2011): Bacteriological quality and histamine-forming bacteria associated with fish meats and environments in HACCP and non-HACCP fish processing factories. Food Cont., 22(10): 1657-1662.

International Commission on Microbiological Specification for Foods “ICMSF” (1978): Microorganisms in foods. Their significance and methods of enumeration.2nd Edi. Toronto, Univ. of Toronto Press 1978.

Jaksic, S.; M.Z.Balos; Z.Mihaljev; P.J.Radulovic and K. Nesic(2017): Comparison of analytical methods for the determination of histamine in reference canned fish samples. Earth Environ. Sci. 85: 012066. 59th International Meat Industry Conference.

Kaper, J.B.; J.P.Nataro and H.L. Mobley (2004): Pathogenic Escherichia coli. Nat Rev Microbiol., 2(2): 123-140.

Korkmaz, C.; Ö.Ay; Y.Ersoysal; M.A.Köroğlu and C.Erdem (2019): Heavy metal levels in muscle tissues of some fish species caught from north-east Mediterranean: Evaluation of their effects on human health. J. Food Comp. Anal., 81: 1-9.

Leszczynska, J.;M. Wiedlochaand U. Pytasz(2004): The histamine content in some samples of food products. Czech J. Food Sci., 22: 81-86.

Lopez Da Silva, M.; G.R.Matte; P.M.L. Germanoand M.H.Matte (2010): Occurrence of pathogenic microorganisms in fish sold in Sao Paola, Brazil. J. Food Saf., 30: 94-110.

Mohammed, A.; H.Abdallah;A.El-Hafez;A. Amin and M. Mousa(2017): Quality assessment of some retailed marine fish and shellfish in Alexandria province. Alex. J. Vet. Scis, 52: 166-72.

Morshdy,  A.M. ; E.A. EldalyandK.I.  Sallam(2002): Sanitary evaluation of some marketed fishes in Sharkia Province. 6th Vet.Zag. Conference: 313-21.

Mustafa, G.; T. Yasmin ; S.S. Nilla; M.R.Khan and A.R. Khan (2013) : Effect of Freezing Period on The Microbial And Nutrient Quality of Pool Barb, PuntiusSophore From Dhaka City Retail Market, Bangladesh. Bangladesh J. Zoology, 41(1): 1-11.

Obemeata, O. ; F.P. NnennaandN. Christopher (2011): Microbiological assessment of stored Tilapia guineesis. African J. Food Sci., 5(4): 242-247.

Popelka, P.; J.Nagy; M.Pipová; S. Marcinčákand L. Lenhardt(2014): Comparison of chemical, microbiological and histological changes in fresh, frozen and double frozen rainbow trout (Oncorhynchusmykiss).ActaVeterinaria, Brino, 83: 157–161.

Prabakaran, P.; K. S.Kannan; M.Anand andV.Pradeepa (2011): Microbiological quality assessment in a fish processing plant at mandapam, ramanathapuram district. Archives App. Sci. Res., 3(2):135-138.

Raafat, M. S. (2004): Some food poisoning agents in shell fish. M.V. Sc., Thesis (Meat Hygiene), Faculty of Vet.Med., Zagazig Univ.

Rohde,  A.;J. A.Hammer; B.Appe ;R. Dieckmannand S. Dahouk(2014): Fishing for bacteria in food- A promising tool for the reliable detection of pathogenic bacteria?. Food Microbiol., 46(1): 395-407.

Shabarinath, S.; H.S.Kumar; R.Kdyhushiramani; I. Karunasagarand I. Karunasagar(2007): Detection and characterization of Salmonella associated with tropical seafood. Int. J. Food Microbiol., 114: 227-223.

Takahashi, H.; M.Dgai; S.Miya; T.Kuda and B.Kimura (2015): Effects of environmental factors on histamine production in the psychrophilic histamine-producing bacterium Photobacteriumiliopiscarium. Food Cont., 52: 39-42.

United States Food and Drug Administration "USFDA" (2001): FDA and EPA safety levels in regulations and guidance: fish and fisheries products hazards and controls guidance (285 p.). WashingtonI: US Food and Drug Administration.

Venugopal, V. (2002): Biosensors in fish production and quality control. Biosensors and Bioelectronics. 17: 147-157.

World Health organization "WHO" (1999): Food safety issues associated with products from aquaculture. Report.Of A joint.World.Health Organ. Tech. Rep. 883, I-vii: 1.

بعض مؤشرات الجودة فی أسماک المیاه العذبة المسوقة

بمحافظة الشرقیة، مصر

علاء الدین محمد مرشدى، عبدالله فکرى عبدالله و اسراء عبدالعزیز

قسم مراقبة الأغذیة –کلیة الطب لبیطرى – جامعة الزقازیق

تعد الأسماک مصدرًا مهمًا للبروتین ذی القیمة البیولوجیة العالیة بالإضافة إلى إستساغتها العالیة وهضمها الجید، فهی تحتوی علی العدید من المعادن الأساسیة والفیتامینات والأحماض الدهنیة غیر المشبعة. تشکل الملوثات البکتیریة و الکیمیائیة خطراً جثیماً على صحة الإنسان حیث أن هذه الملوثات تتراکم فی الأسماک بمستویات یمکن أن تسبب أضرار متعددة علی صحة المستهلکین. لذلک أجریت هذه الدراسة لتقییم بعض مؤشرات الجودة على بعض

الأسماک فتم تجمیع عدد 100 عینة عشوائیة من الأسماک (أسماک البلطی النیلی والقرموط) بواقع 50عینة لکل منها من الأسواق المختلفة فی مدینة الزقازیق، محافظة الشرقیة، مصر. أوضحت نتائج الفحص الظاهری لعینات أسماک البلطی النیلی والقرموط أنها مقبولة وصالحة للإستهلاک الأدمی بنسبة کلیة 94,2% و 90,7% لکل منهما علی الترتیب.أوضحت نتائج الفحص البکتیرى أن متوسط العدد الکلى للبکتیریا الهوائیة ​​کان6,63± 0,06 و 6,58 ±0,057 لوغاریتم 10 مستعمرةبکتیریةلکلجرامفیأسماک البلطی النیلی والقرموط و العدد الکلى للبکتیریا المعویة تراوحت قیمته من 3,81 إلی 5,52 لوغاریتم 10 مستعمرة بکتیریة لکل جرام فی أسماک البلطی النیلی، بینما تراوحت قیمته من 2,7 إلی 5,57 لوغاریتم 10 مستعمرة بکتیریة لکل جرام فی أسماک القرموط.أوضحت نتائج الفحص الکیمیائی أن 90% من أسماک البلطی النیلی إحتوت علی الهیستامین، بینما إحتوت کل عینات القرموط علیه. تراوحت قیمة الهیستامین من 1 إلی 22,7 و 1,2 إلی 31,9 مجم / 100جم فی کل من أسماک البلطی النیلی والقرموط.لذلک توصی هذه الدراسةبالمعالجة الصحیة للأسماک بدایة من الصید إلى نقطة الاستهلاک و سرعة تبرید أو تجمید الأسماک والحفاظ على درجة حرارتها منخفضة حتى وقت الاستهلاک بالإضافة إلی معاملة الأسماک حراریاً عند درجات حرارة مناسبة قبل الإستهلاک.