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Storage conditions and their impact on the sanitary quality of stored food.

INTRODUCTION

Depreciations of stored foodstuffs are generally the result of pest activity, inadequate storage mode or type of commodity that can be a very responsive cultivar to loss factors [43]. The same author reports that over 30% of production is lost between harvest and consumption. Storage is not a neutral operation. If the storage system has seen a great improvement on a large scale, it remained, however, mediocre among farmers, grain merchant and households especially in precarious habitats that are of manifest existence in the suburbs and even in urban centers cities in the world and in Algeria. The precarious habitats are a real factor of vulnerability of the population. They are mainly built on land as the banks of arroyos and canals, or marshy land or unhealthy. This implies that the storage conditions are far from being met and that these commodities are directly subject to the vagaries of conditions in these environments. These conditions are, however, a favorable environment for the development of storage pests. In farmers' attics where more than 80% of crops are stored, an important part of these stocks can be destroyed by pests [33,16,42]. Corn grain losses due to weevils can exceed 25% of the crop or even 40% in six months of storage [59] & Papachristos Stamopoulos, [46]. These pests are diverse (insects, mites, and fungi) [53] and Chambers [12], and their impact is not limited only to physical and chemical degradation products as it was reported by Domngang, [15] and Kamdem, [32], but they are the cause of many diseases. Bennett and Klich [6] noted That Among the hundred mycotoxins approbation at present, thirty is really significant for human and animal health due to Their frequency gold Their toxicity.

One of the most serious problems to confront the quality of rice is the presence of mycotoxins which are produced by different species of the genus Aspergillus or Penicillium. A flatoxins are of greatest concern As They are highly toxic, mutagenic, teratogenic and carcinogenic compounds That-have-been Implicated as causative agents in human hepatic and extrahepatic carcinogenesis [10,26,27,28,38]. According to Mehrez [40] mycotoxins that are prouced can remain on food stuffs and transfer throughout the food chain in derivatives. Some studies-have found the implication of ochratoxin A in some epidemic nephropathy in animals and humans [6,11,49]. According to Guerre [21], since the 1960s, aflatoxicosis were recorded and reproduced experimentally on a large number of animal species. According to the same author, retrospective studies have permitted to suspect the intervention of these molds in a number of clinical manifestations of etiology unknown till now. According to Nguyen [43] an investigation of liver or kidney cancer diseases is recommended to evaluate the impact of mycotoxin contamination on the health of the population. Our study, however, focuses on the inventory of pests that can develop in foodstuffs stored in ambient conditions and through a test showed the denaturing effect caused by these pests on health. The experiment was conducted on mice that underwent various tests

This study is to conduct a test in the laboratory on mice that are subject to a special diet. After a month of rearing under controlled conditions and the food is just basic denatured foods obtained itself after one year of storage at ambient conditions. The tests carried out are to highlight the impact of foods affected by pests on human and animal health.

MATERIALS AND METHODS

Food stuffs which have the subject of this study were purchased from a grain merchant in the region of Algiers. The foodstuffs were intended for human consumption The different products are: Maize n = 06 and, Wheat n = 06 Both types of products, divided into 12 batchs with a weight of 500 g, were placed in glass jars and are stored in home storage conditions (humidity and ambient temperature).

Gqaleni et al. [19] noted that water activity (aw) and temperature are two of the major environmental most factoring to control development of fungi and to affect their metabolic activities.

After a year of storage, stored products (a sample of each batch) were the subject of a fungal, entomological and acarological analysis. An inventory of pests (insects and mites) was established as well as a list of fungal populations.

2.1. Methodologies:

2.1.1 Entomological analysis:

A sample of each product was subject to direct observation under a binocular magnifying glass. A list of all insects' species was recorded after determining.

2.1.2 Acarological analysis:

The study of the acarological fauna is conducted using the Berlese technique. The species found are determined according to determination keys.

2.1.3 Isolation and identification of fungal accompanying interior on the seeds:

The seeds were sterilized with 10% sodium hypochlorite solution for 2 min and rinsed twice with sterilized distilled water, the seeds were dried with sterile filter paper and plated (5 seeds per plate),on fresh Potato Dextrose Agar(PDA) medium impregnated with (10% of Acetic Acid solution, 1ml /100ml of medium), each replicate has three plates, and incubated for 7-10 days at 27[degrees]C. The resulting of borne fungus colonies on seeds were sub-cultured by transferring a small mycelia plugs from the colony margins. Pure culture was obtained by sub-culturing three times [7]. The fungi were identified on the basis of their morphological and reproductive characters [7,54]. All samples were identified in Laboratory of Microbiology, Department of Botanic, National Higher School of Agronomy (Algeria), and verified by Professor Keddad, Department of Botanic, National Higher School of Agronomy, Algeria.

2.1.4. Biological material:

Tests are performed on male white mice of the species BALB / c from the Pasteur

Institute, Algiers. These mice having a weight of 20 g, were placed in transparent plastic cages of 55 cm long, 33 cm wide and 19 cm high.

The mice are divided into 4 groups, each containing 05 mice:

Two witness groups fed for a month on sane maize and wheat.

Two groups fed for a month on denatured maize and wheat.

The breeding of these animals was performed in a middle space (L = 6 m, w = 3.15 m, h = 2.20 m), the construction of the walls and floor facilitate cleaning and disinfection The mice were divided into 4 experimental groups' of5 mice each. The animals are placed together in polypropylene cages (32cmx2Ocmxl3cm) at a temperature of 24 [degrees] C with humidity of 50%. The food and water are given ad libitum. The litter is renewed every 06 days.

--Tests conducted on mice After 30 days of experiment:

--In calculating the weight gain:

The weight gain is defined as the difference between the final and initial weight of mice (before and after experimenting).

--At the hepato-somatic index (IHS)

The hepato-somatic index is calculated from the ratio between the weight of the liver and the body, multiplied by one hundred.

This index provides information on the state of liver hypertrophy [34]. For this study, this study, all animals were sacrificed and the kidneys and livers are removed and weighed fresh immediately after dissection to avoid drying.

After weighing; these organs were preserved in a liquid strong aqueous Bouin for histopathological examinations.

2.1.5 Histological Part:

Histological sections were performed only on the livers and the kidneys, after a month of diet ingestion. The technique used is that proposed by Martoja and Martoja [37] with hematoxylin / eosin. These histological sections are performed in the laboratory of anatomopathology of Pasteur Institute, Algiers.

Results:

3.1. Inventory of different pests:

The inventory of species of insects collected in stored products which were the subject of this study revealed the presence of 14 species whose numbers vary from one species to another and from one commodity to 'other. Some species show a presence at all such foods is the case of, Mayetiola destructor, Cecidomya sp.1, Oryzaephilus surinamensis, Rhizpopertha dominican and Tribolium confusum. Others marked their presence at the level of a single commodity such as Tribolium castaneum, Oryzaephilus mectator, Bradysia sp., Agromyzidae sp., and Aulacurthum solani, on maize and Cecidomya sp2, Sitophilus oryzae, Nemapogon granella, and Conwentzia hageni on wheat.

The inventory of species of mites in stored products, revealed the presence of 5 species of which three are species that are subservient to stored products such Tyrophagus putrescentiae, Lepidoglyphus destructor, Glycophagus domesticus while Acarus siro is a cosmopolitan species that can be found everywhere and Peymotes sp. which is a predatory species of Bruchidea.

Fungi inventoried: Aspergillus, Rhizopus, Penicillium, Fusarium, Cladosporium, Humicola

Fungi are particularly strongly represented in the maize where 05species were recorded whereas we noted only 03 species. in wheat species Aspergillus and Penicellum mark their presence in Maize as well as in wheat

Fusarum marks his presence in the Maize only

3.2. Test On Mice:

3.2.1. Weight Measurement:
Table 1: Weight gain in mice

              Wheat    Maize

              Body weight (g)

Witnesses     20       20
Processed     27,5     27,66
Weight gain   7,5      7,66


The measuremets relative to body weights of the mice on the test reveal that the contribution of denatured wheat as a nutritional support has a positive effect on gaining weight in these guinea pigs. Indeed the average resulting surplus is of 7.5 grams. There were the same remarks for the batch fed on infected corn. The recorded surplus is of 7.76 grams

The measurements of the relative mass of organs ie kidney and liver of mice on the test showed a decrease in weight and this for both types of food and at the two organs.

The group of mice fed on wheat showed a drop in weight of 0.08 for the liver and of 0.04g for the kidney. Yet for those mice fed on maize, the loss is 0.07g for the liver and 0.05g for the kidney.

3.2.2. Hepato-somatic index:

Hepato-somatic index in mice fed on infested wheat 1.32 / 27.5 * 100 = 4.81

Hepato-somatic index in mice fed on sane wheat 1.4 / 28.8 * 100 = 5.08

Hepato-somatic index in mice fed on infested maze 0.99 / 27.66 * 100 = 3.58

Hepato-somatic index in mice fed on sane maze 1.06 / 27 * 100 = 3.93

Measuremenst of liver somatic index performed on the mice that underwent experiment have shown a decreasing trend of this index in mice fed on wheat as as in mice fed on maize. The recorded values in mice fed on undamaged wheat 05.08 while for the contaminated wheat the value is 04.81. as for corn, the value this sound food is 3.93 while for the infected products, it is 3.58

3.2.3. Microscopic observation:

The report of the general autopsy carried out on the batch of mice fed on sane maize showed no anomalies in both the liver or kidney by contrast the group of mice fed contaminated maize, there was the presence in the liver, a yellowish lesion to orange yellowish lesion noticeable on the surface and the cut. In the kidney no abnormalities were reported.

The group of mice fed on sane wheat shows nothing wrong in the kidneys and livers. Whereas the mice fed on contaminated wheat, have yellowish lesions and reddish lesions visible puncture on surface and cut in the liver and nothing in the kidneys. The main lesions seen during the microscopic examination of liver are yellowish and reddish hepatic infiltrations are probably degenerative from alimentary or inflammatory origin

3.2.4. Histopathogycal exam:

Histopathological examination carried out on the batch of mice undergoing contaminated nourishment based on maze highlighted the presence of hyperplasia of bile ducts, a liver steatosis and vascular congestion. This histopathological lesion was compatible with mycosis toxicosis (contamination of food by mycotoxins).

The presence of a moderate surplus fat and glycogen was noted in the liver. This steatosis and hepatic glycogen are frequently observed lesions in the case of a diet high in carbohydrates.

The batch of mice with a sane diet has presented an overcharge lipid and liver glycogen minimal.

Examination of the batch of mice having undergone a contaminated wheat-based diet showed the presence of parenchymal inflammatory infiltration and minimal perivascular multifocal and minimal fat overcharge the liver.

The batch of mice with a diet of sane wheat highlights the presence of inflammation peri-portal.

Discussion:

Following this study, the inventory revealed the presence of 18 species in six orders that are Diptera, Coleoptera, Lepidoptera, Hemiptera, Nevroptera and the Mites. By order of species richness we found many works that refer to the attack and the loss of stocks of cereals and legumes because of insects [60,52,3,35] mentioned that all the stored products are under attack from many insects belonging to the orders Isoptera, Hemiptera, Orthoptera, Lepidoptera and Coleoptera. These species, in addition to the damage, they cause in grains or foods, are vectors of mold spores they introduce in the interior of the same grain through lesions they create. The contamination of peanut, cotton, corn with A. flavus and aflatoxin before harvest is often linked to the attack by insects.

Since the initial era when man began to cultivate cereals and store food, deterioration by molds is inevitable. The food is gradually invaded by a fine down (duvet) (mycelium) white, black, green, orange, red and brown. These molds acidify, fade, ferment and make these unpleasant or even dangerous products [51]. According to Hussein and Brazil [26], the worldwide contamination of foods and feeds with mycotoxins is a significant problem. According to the same author, the economic impact of mycotoxins includes loss of human and animal life. Increased health care and veterinary care costs, reduced livestock production.

Mycotoxins are secondary metabolites of low molecular weight, present in many products of human and animal food and which cause many diseases in humans and animals [39,13]. Thousands of toxic molecules have been identified in fungi but only twenty families set problems in human or animal nutrition [9]. In fact, the results obtained in this study revealed a number of abnormalities in kidneys and livers of the mice in the experiment.

4.1 Weight gain:

The results for weight gain showed a positive effect. Indeed, mice fed denaturated food took over weight which is 7.5 grams for the batch fed on denatured wheat 7.76 gram for the batch fed on denaturated maze. Guerre et al [21] noted a loss. However it was found that the organs, in this case the kidney and liver of the experiment, have lost weight on the contrary to the work of Gomes and et al. [20] noted an increase in liver size which probably explains a weight gain.

4.2 Hepato-somatic index:

Measurements of hepato-somatic index performed on the mice that underwent experiment have shown a decreasing trend of this index in the mice fed on wheat well as on maze. The recorded values in mice fed on sane wheat are 5.08 while contaminated wheat the value is 4.81. As for corn, the value for the sane maize 3.93 while for the infected product, it is 3.58. These results seem to confirm the malfunction of the livers of mice that have undergone a diet containing mycotoxins

4.3 Microscopic Observation:

Microscopic observation revealed the presence of a lesion visible yellow orange yellow surface and cutting of mice the liver nourishes based but contaminated. Kidney no abnormalities were reported. In mice fed on contaminated wheat, there the presence of yellowish lesions and reddish lesions visible puncture surface in the liver and not the kidneys. These lesions seen during the microscopic examination of liver are yellowish and reddish infiltrations are probably degenerative or inflammatory food.

4.4 Histopathological examination:

This examination revealed abnormalities in the liver and in the kidney.

4.4.1 At the kidney level:

Renal congestion has been demonstrated that, according Jeffrey et al. [31] is a cause of renal dysfunction. It was shown that partial occlusion of the renal vein leads to an immediate reduction in renal blood flow congestion. This resulted in a significantly more complex situation than that observed in an animal with renal vein partially occluded. Physiology seems to be more complicated than a simple hydraulic effect causing decreased perfusion pressure.

The numerous works carried out around the world have shown that mycotoxins have greater effects on the kidneys. In fact, Pfohl-Leszkowicz et al., [49], noted that in 1950, a series of publications describing renal disease in Bulgaria and Romania, which has become like a Balkan endemic nephropathy (BEN). This pathology was characterized by tubular degeneration, interstitial fibrosis and hyalinization of glomeruli accompanied by enzymuria and impaired renal function without nephrotic syndrome. Later, an association between nephropathie endemique and tumours of the kidney pelvis and ureter was recognized, so that the problem of BEN became not only nephrological, but also oncological. There may also be an association with increased urinary bladder cancer incidence, although many confounding factors may interfere in the analysis of data for this organ. The factors that are responsible for this pathology are varied but the way of fungal toxins is most suspected. In a biostatistical study of urinary tract tumors conducted later by Nikolov et al. [45], confirmed the positive correlation between the incidence of urinary tract tumors (UTT) and BEN demonstrated in our first population-based case control 1977 study and that the percentage of food samples and blood containing nephrotoxic mycotoxin and carcinogen ochratoxin A (OTA) correlated with the origin of echantillonsr Nikolo (the group of BEN/UTT patients). The most contaminated samples were found in BEN villages and households, and the urinary excretion of OTA was higher in the group of BEN/UTT patients. The same results are obtained by Peraica and et al. [48] and Mally et al., [36] who noted that Ochratoxin A (OTA) is an ubiquitous nephrotoxic and carcinogenic mycotoxin considered to be involved in the aetiology of Balkan endemic nephropathy (BEN).-Ochratoxin A is toxic to humans and animals. It is potentially nephrotoxic in all species tested, with the exception of mature ruminants. OTA is considered a renal carcinogen at least during a long exposure. It was classified in Group 2B. It causes, for example, various morphological abnormalities in the rat, mouse, hamster, pig and chicken embryo. OTA affects cellular and humoral immunity. Peraica and et al. [47] noted that the most imputed causative agent of endemic nephropathy is the mycotoxin ochratoxin A (OTA), because its confirmed nephrotoxic and carcinogenic action.

4.4.2 At the liver level:

Histopathological examination carried out on the batch of mice fed contaminated maze highlighted the presence of hyperplasia of bile ducts, a liver steatosis and a hepatic vascular congestion. These histopathological lesions were compatible with mycosis toxicosis (contamination of food by mycotoxins. According to Benedict and et al. [5], bile duct hyperplasia is liver precancerous lesions Hepatic vascular congestion occurs when there is an excessive consumption or the presence of toxins in the blood that cause liver dysfunction.

The same examination revealed the presence of a yellow to orange yellow lesions visible on the surface and cut in the liver. According to Guerre, the aflatoxicoses are characterized in their forms acute and sub-acute by the appearance of toxic hepatitis with cytolysis and proliferation of bile ducts and the effects of aflatoxin on animal health vary according to the species. The AFB1 causes hepatotoxicity, it is teratogenic and immunotoxic. To be toxic to humans, suggestions of a relationship between in AFs heavily contaminated foods and some diseases are more recent. Indeed acute poisoning, potentially linked to consumption of FAs, was identified by Hall & Wild [22] and Wild & Hall [61]. Contamination citrinin is observed in various grain-based foods (corn, wheat, barley, rice, fruits, grain products ...) [30,14,1,4,41]. Angelita [2] has noted that the clinicopathological aspects aflatoxicosis cases in dogs are described in southern Rio Grande do Sul. Indeed, with a total of 27 dogs with liver cirrhosis, six on suspicion of aflatoxicosis characterized by macro and microscopic lesions. The macroscopic changes were characterized by ascites, by jaundice, and increased liver size, with or without nodules, as well as bleeding. These cases were classified according to the major histological appearance characterized by diffuse vacuolation in the cytoplasm of hepatocytes in acute cases, proliferation of the bile ducts and mild fibroplasia in subacute cases and severe fibrosis in chronic cases Guerrre et al., [21] suspect the intervention of these molds in a large number of clinical etiologies of previously unknown.

The acute form of intoxication was described by Shank [57] and Shank [58]. It is similar to events observed in most experimental reproduction of the disease in many animal species (domestic mammals, laboratory rodents, birds, fish) and is characterized by hepatocyte necrosis lesions with proliferation of bile canaliculi.

According Rocan et al., [55], aflatoxicosis is suspected in Reye's syndrome in children due to neighboring clinical manifestations of those observed during aflatoxicosis in monkeys as it was reported by Bourgeois [8] (vomiting, seizures, coma associated with fatty infiltration of the liver, kidney and heart and a brain edema. the same author association between liver cancer, hepatitis B and aflatoxin exposure is a serious problem that has to the foundation hepatocellular carcinogenicity of aflatoxin B1 (AFBI) animal and interpretation of numerous epidemiological studies that a high prevalence of liver cancer can be associated with high contamination of the food environment in aflatoxins.

Conclusion:

The results obtained show that pests in stored food stuffs are a serious problem to both the human and animal health. Their presence is accompanied by an establishment of molds that are at the origin of synthesis of mycotoxins. These have proven highly toxic and even carcinogenic. Indeed the risk evaluation of fungal contamination of man's and animals' food is confirmed by numerous studies.

However it requires, firstly identifying the species which may contaminate substrates and to determine whether, in the preparation conditions, environmental conditions can result in the synthesis and accumulation of toxins in food. This work is a prerequisite for the establishment of relevant mycotoxic control plans, taking into account the peculiarities of the different foods.

These results are from new data when the effect of mycotoxins on human and animal health. This creates a tight control when storing food particularly in too humid and for all coastal regions.

Prospects:

Despite all the work done around the world, the results of this work might be more than an extra alert on the contamination of stored food with mycotoxins in warmer areas and where the humidity is quite high.

Food storage must be done in good conditions in order to avoid mildew.

These studies open new perspectives when looking for new methods of food storage and preservation in better conditions.

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(1) MAHDI Khadidja, (2) BENLAMEUR Zahia, (2) GHEZALI Djenoul and (2) BELHAMACHA Mounira

(1) Agronomy department, faculty of sciences of nature and life and terresteral sciences, University ofBouira, Algeria

(2) Departement. of Agricole Zoology and Forestry National Higher Agronomic School, El-Harrach, Algiers, Algeria

Address For Correspondence:

MAHDI Khadidja, Agronomy department, faculty of sciences of nature and life and terresteral sciences, University of Bouira, Algeria Tel: +213550373460; E-mail : mahdi.khadidja@yahoo.fr

This work is licensed under the Creative Commons Attribution International License (CC BY).

http://creativecommons.org/licenses/by/4.0/

Received 22 March 2016; Accepted 28 May 2016; Available online 12 June 2016
Table 2: Relative mass weights (liver and kidney)
of mice fed on wheat and maiz

           Liver             Kidney

           wheat    mais     wheat    mais

Temoins    1,4      1,06     0,45     0,4
Traites    1,32     0,99     0,41     0,35
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Author:Mahdi, Khadidja; Benlameur, Zahia; Ghezali, Djenoul; Belhamacha, Mounira
Publication:Advances in Environmental Biology
Article Type:Report
Geographic Code:6ALGE
Date:May 1, 2016
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