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1 IJA 1186.doc 1 2 REVIEWER’S REPORT 3 General Comments 4 Generally, while the study offers some fairly good information about types of farmed shrimp diseases in some ‘shrimp growing provinces’ in India, it does very little to help the reader conceptualize the spread of the diseases and the enormity of the problem. 5 6 7 8 9 10 11 12 13 14 The abstract is dry and tells nothing about where, when and how study was conducted. If, on the other hand, the report was intended to only catalogue the diseases of farmed White Leg Shrimp in India, then the reader ought to, at least, be given the benefit of the etiology of the diseases. There is virtually no methodology. The method of study must be clearly spelt out and for such a study, a description of the study area including maps is appropriate. 17 The pictorial representation of the shrimp diseases may be generally acceptable but it must be used to illustrate a thorough description. Most of the pictures are, nonetheless, unsatisfactory. 18 The discussion is generally acceptable. 19 There is no conclusion. The two sentences made almost as part of the discussion cannot pass for conclusion. 15 16 20 21 22 23 24 25 26 27 28 English language has not been succinctly applied in this article and the author is urged to take note of this during the revision. Colloquial English is not permitted in scientific writing. Also, such phrases or terms as ‘in a nutshell’ must be avoided at all cost. I have attempted to put in some corrections on the main manuscript to serve as a guide for future revision of the article. The article, as it stands presently, adds very little or no knowledge to science and therefore cannot pass for publication in a reputable scientific journal. 2 29 30 Diseases of White Leg Shrimp Litopenaeus vannamei Culture in India 31 32 33 34 35 36 37 38 39 *Gunalan B and Soundarapandian P CAS in Marine Biology, Faculty of Marine Sciences, Annamalai University, Parangipettai, Tamilnadu, India 40 41 42 43 44 45 46 47 48 49 50 51 *Corresponding Author : B.Gunalan, Center of Advanced Study in Marine Biology, Faculty of Marine Sciences, Annamalai University, Parangipettai-608 502, Tamil Nadu, India, Tel: (Off): 04144- 243223 and Fax number 04144- 243553, e-mail: [email protected] 3 52 Abstract 53 The knowledge about disease is very much essential to keep the animal in disease free 54 environment. In the present study eight diseases were reported in the culture ponds of L. 55 vannamei including Black gill disease – Andrapradesh, IHHNV-Andrapradesh and Orissa, 56 WMD -Tamilnadu and Andrapradesh, WSSV -Andrapradesh. White gut disease - Tamilnadu and 57 Andrapradesh, Muscle cramp disease- Tamilnadu and Andrapradesh. The symptoms of each 58 disease and their possible cure were also described in details. The outbreak of disease in shrimp 59 culture definitely affects profit, from a farmer’s point of view, and deterioration of flesh quality, 60 from a consumer’s view point . Many diseases are already reported in P. monodon in India, 61 however, no such diseases are reported for L. vannamei a. So study on diseases is very important 62 to create awareness among farmers to prevent the diseases. In a nutshell, farmers should stock 63 healthy disease free seeds, at optimum stocking densities, maintain good water quality and 64 provide nutritious feed for higher survival and growth. 65 Keywords: Litopenaeus vannamei; Black gill disease; White gut disease; IHHNV; Muscle 66 cramp syndrome; White leg shrimp disease; Shrimp culture 67 68 Introduction 69 Shrimp farming is a multi-billion dollar industry contributing a major income to several 70 countries in Asia and South America. The rapid growth of shrimp farming led to an economic 71 boom but, unfortunately, the outbreak of viral diseases has increased the economic risks and 72 slowed the industry’s development (Flegel, 2006). The most important diseases of cultured 73 penaeid shrimp, in terms of economic impact, in Asia, the Indo-Pacific, and the Americas have 4 74 infectious etiologies. Among the infectious diseases of cultured shrimp, certain virus-caused 75 diseases stand out as the most significant. The pandemics due to the penaeid viruses WSSV 76 (White spot Syndrome Virus) and TSV (Taura Syndrome Virus), and to a lesser extent IHHNV 77 (Infectious Hypodermal and Hematopoietic Necrosis Virus) and YHV (Yellow Head Virus), 78 have cost the penaeid shrimp industry billions of dollars in lost crops, jobs and export revenue. 79 The social and economic impacts of the pandemics caused by these pathogens have been 80 profound in countries in which shrimp farming constitutes a significant industry. In the present 81 study some of the common diseases in L. vannamei were recorded all along the state of 82 Andrapradesh, Orissa and Tamilnadu and described in detail. 83 Material and methods 84 During the study period diseased L. vannamei were collected from different shrimp 85 farms located in the state of Andrapradesh, Orissa and Tamilnadu. Whole shrimps were 86 immediately immersed in 95% ethanol in plastic containers. Samples were maintained at 40C 87 until processing. 88 89 Results and Discussion 90 Black gill disease 91 In the present study black gill disease was observed at shrimp farms in Andrapradesh. 92 The black gills disease is very common in penaeid shrimps. It often occurs when there is too 93 much plankton in the water or too much mud on the pond bottom. It is especially a problem in 94 ponds with high population density (over 60 shrimp/m2), insufficient aeration or when the water 5 95 is not changed frequently enough or irregular probiotic application. Before the shrimp appear 96 diseased or start to die its gills turn black. 97 98 99 100 Fig.1 Black gill disease 101 Black gill disease is reasonably easy to cure. If farmer noticed dark-colored gills in some 102 shrimp it is suggested to change the water more often and also provide more aerators. This will 103 usually solve the problem quickly. Addition of lime according to the pH concentration is also 104 recommended. Similar disease was already reported by Lightner and Redman, (1998), 105 MPEDA/NACA (2003), Doughtie and Rao (1983) and Johnson (1995) (Fig 1& 2). 6 106 107 108 109 110 111 112 113 Fig.2 Black gill disease observed during sampling IHHNV (Infectious Hypodermal and Hematopoietic Necrosis Virus) 114 In the present study, IHHNV (Infectious Hypodermal and Hematopoietic Necrosis Virus) 115 disease was observed at shrimp farms in Andrapradesh and Orissa. IHHNV is very common in 116 grow out ponds, especially when the postlarvae brought from broodstock that were raised in 117 earthen ponds in Thailand. The white leg shrimps with IHHNV are easy to observe because they 7 118 display a typical clinical signs of a deformed or stunted rostrum (Fig. 3). It might be bent to one 119 side and the body of the shrimp might be crooked. These clinical signs will be apparent after the 120 shrimp have been kept in grow out pond for 30 days. 121 122 Fig.3 IHHNV (Infectious Hypodermal and Hematopoietic Necrosis Virus) affected shrimps 123 In general the deformed shrimp will be seen in only 10-20% of the shrimp population, but 124 if it is very severe, 70-80% will be affected. The diseased shrimp will grow slowly and will have 125 a low survival rate, so the overall yield will be lower. However, the diseased shrimp will not 126 usually die, they will just be weakened. The occurrence of IHHNV is likely to rise in the future 127 as more farmers use post larvae produced from brood stock from local shrimp farms than 8 128 imported ones. Outbreaks of IHHNV can occur at any time of the year. The main risk factor is 129 the source of the shrimp larvae. It is very unlikely that larvae produced from specific pathogen- 130 free parents will get IHHNV. 131 The virus which causes infectious hypodermal and haematopoietic necrosis infects many 132 species of shrimps including P. monodon, P. japonicus, P. stylirostris, P. vannamei, 133 P.semisulcatus, P. occidentalis and P. californiensis. IHHN has reported in central and South 134 America, India, Indonesia, China, Taiwan, Thailand, Malaysia, Philippines, Hawaii, Guam, 135 French Polynesia, New Caledonia and Israel ( Bondad-Reantaso et al., 2001). The disease IHHN, 136 and later its causative agent, IHHNV, was first described as the cause of acute epizootics and 137 mass mortalities (> 90%) in juvenile and subadult of L. stylirostris farmed in super-intensive 138 raceway systems in Hawaii (Brock et al. 1983; Lightner, 1983, 1988; Lightner et al. 1983, 139 1983a; Brock and Lightner, 1990). Shortly after its discovery in L. stylirostris, the virus was 140 found in L. vannamei being cultured at the same facility in Hawaii and these L. vannamei were 141 shown to be asymptomatic carriers of the virus (Lightner et al, 1983a; Bell and Lightner, 1984). 142 Hence, the economic and production impacts of IHHNV infection in L. vannamei are due to 143 reduced and irregular growth and small sized shrimp at harvest and not elevated mortality. To 144 mitigate this effect, several strategies have been used. With one strategy, selected lines of L. 145 stylirostris, which were not only resistant to IHHN disease, but are also refractory to infection, 146 were developed (Dhar et al., 2001). IHHNV-free lines of L. vannamei were also developed as 147 SPF (specific pathogen-free) lines and these stocks were the first developed in the SPF stock 148 development program (Pruder et al., 1995). 149 White muscle disease 9 150 In the present study WMD (White muscle disease) was observed at shrimp farms in 151 Tamilnadu and Andrapradesh. The epizootic of WMD was first recorded in November 2001 in 152 few hatchery reared PL of M. rosenbergii in Andrapradesh and Tamilnadu states. More than 18 153 cases of WMD in freshwater prawn hatcheries with PL mortalities ranging from 30% to 100% 154 were recorded from November 2001 to December 2002. The first sign related to this disease was 155 the poor feeding (mineral deficiency in water, poor quality feed) and lethargy of the prawns 156 especially during the first 5 days of PL settlement resulting in slow mortality (Fig. 4 &5 ) 157 158 Fig.4 White muscle disease in bigger size L. vannamei 10 159 As the morphological and clinical signs of the disease reported by Chen et al. (2001) 160 were similar to the WMD recorded in this study. It is likely that the involvement of 161 162 Fig.5 White muscle disease in small size of L. vannamei 163 The Gram-positive cocci, Lactococcus garvieae in WMD found by those workers could possibly 164 have been a secondary infection. The disease progressively destroyed the abdominal muscular 165 organization of the prawns especially the striated muscles finally leading to mortality. Several 166 authors reported white muscle disease in shrimps and prawns (Chen et al., 1992; Sudhakaran et 167 al, 2006; Ravi et al ., 2009). Besides the above, this white muscle disease also recorded in L. 168 vannamei in the present study. 169 White gut disease (WGD) 170 In the present study white gut disease observed in L. vannamei shrimp farms from 171 Tamilnadu and Andrapradesh. Vibriosis is one of the major disease problems in shellfish and 172 finfish aquaculture. Vibriosis is a bacterial disease responsible for mortality of cultured shrimp 173 worldwide (Lightner and Lewis, 1975; Adams, 1991; Lightner et al., 1992; Lavilla-Pitogo et al., 11 174 1996; Lavilla-Pitogo et al., 1998; Chen et al., 2000). Vibrio species are widely distributed in 175 culture facilities throughout the world. Vibrio-related infections frequently occur in hatcheries, 176 but epizootics also commonly occur in pond reared shrimp species. Vibriosis is caused by gram- 177 negative bacteria in the family Vibrionaceae. Outbreaks may occur when environmental factors 178 trigger the rapid multiplication of bacteria already tolerated at low levels within shrimp blood 179 (Sizemore and Davis, 1985), or by bacterial penetration of host barriers. The exoskeleton 180 provides an effective physical barrier to pathogens trying to penetrate the external surface of 181 crustaceans, as well as the foregut and hindgut (Fig 7a & b). 182 183 (7 a) Fig. 7a & b. White gut disease in L. vannamei (7 b) 184 The occurrence of five types of diseases: tail necrosis, shell disease, red disease, loose 185 shell syndrome (LSS) and white gut disease (WGD) is by Vibrio spp. in P. monodon from 186 culture ponds of coastal AndhraPradesh (Jayasree, et al., 2006). Among these, LSS, WGD, and 187 red disease caused mass mortalities in shrimp culture ponds. Six species of Vibrio—V. harveyi, 188 V. parahaemolyticus, V. alginolyticus, V. anguillarum, V. vulnificus and V. splendidus—are 189 associated with the diseased shrimp. The distribution and species composition of luminous 190 bacteria in commercial penaeid shrimp hatcheries were studied by Jawahar Abraham and 12 191 Palaniappan (2004). The observation on the presence of V. harveyi (97.30%) and V. orientalis 192 (2.70%) in shrimp gut contents evinced that the primary source of these bacteria in a shrimp 193 hatchery was the faecal matter from brood stock, possibly at the time of spawning. Mortalities 194 due to vibriosis occur when shrimps are stressed by factors such as: poor water quality, 195 crowding, high water temperature, low DO and low water exchange (Lewis, 1973; Lightner and 196 Lewis, 1975; Brock and Lightner, 1990). 197 Muscle cramp syndrome (MCS) 198 In the present study muscle cramp disease observed in L. vannamei shrimp farms of 199 Tamilnadu and Andrapradesh. The temperature fluctuations affect the muscle of the shrimps. 200 Muscle cramp disease is very common in grow out ponds (Fig.8). It is attributed that less amount 201 of dissolved oxygen leads to mass mortality in ponds (Fig. 9). This is very common in high 202 stocking ponds. Once DO level was reduced leading to stress among the shrimps, finally they 203 will die especially inside and sides of the culture ponds. 204 205 Fig. 8 Muscle cramp syndrome 13 206 207 Fig.9 Mass mortality in the check tray due to dissolved oxygen depletion 208 The seven diseases mentioned above are serious problems for raising white leg shrimp in 209 intensive culture systems. The farmers should have basic knowledge about these diseases and 210 their prevention. If the farmers stock healthy disease free seed and maintain good water quality 211 and provide nutritious feed to get higher survival. 212 References 213 214 Adams, A., 1991, Response of penaeid shrimp to exposure to Vibrio species. 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