Preserving young‐of‐the‐year Perca fluviatilis in ethanol,formalin, or in a frozen state and the consequences for measuring morphometrics

As it is often not possible to immediately analyse individuals sampled in the field, captured fish are preserved and stored for later investigation. The objective of this study was to assess the effects of various preservation methods on subsequent changes in morphometric parameters while also providing correction factors to re‐calculate the original body dimensions when sampled fish are measured at a later date. In this study, juvenile perch (Perca fluviatilis, 66.5 ± 10.2 mm total length) were measured directly after capture, then either frozen at ?20°C, preserved in 70% ethanol, or in 4% formalin. They were again measured after 24 h, 3 days, 7 days, and thereafter on a weekly basis for 8 weeks. Ethanol‐preserved perch greatly decreased length and weight; formalin preservation also led to a comparable length reduction, but increased the weight of the perch. In contrast, frozen perch showed less shrinkage and only moderate weight loss. Of the three preservation methods, freezing clearly caused the fewest distortions. Hence, freezing is recommend as the most preferable preservation method, especially in multi‐disciplinary studies on fish ecology; for all other preservation methods the correction factor over time will have to be pre‐determined for each species and size class.     

Effect of conservation of fish in formalin and ethanol on length‐weight relationships and condition factor in Tlaloc labialis (Günther, 1866)

The effect of the preserver on the length‐weight relationship and condition factor were estimated using 50 specimens of Tlaloc labialis; a small stream fish from the Grijalva River basin, México. They were measured at different stages of preservation: freshly collected, fixed in 10% formalin and preserved in 70% ethanol for over a 2 years period. All fish decreased in weight and condition factor, with very significant differences compared to the fresh material. The value of the coefficient (b) increased and stabilized from the second month of preservation in ethanol.     

Accuracy of body mass estimates of formalin-preserved fish – a review

This paper highlights possible effects of physical and chemical mechanisms of formalin fixation and preservation on biological tissue and reviews the consequent potential inaccuracies on estimates of body mass of small fishes fixed and preserved in formalin. Twenty-six papers including 65 independent experiments with 35 species which examine effects of formalin on body mass estimates on small fishes are included. The effect of the formalin on the specimens depends on the salinity of the water used to dilute the commercial formalin (usually 1:9 formalin: water) before being used to fixate and preserve fish. Mean wet body mass of the specimens from the studies using seawater or fresh water diluted formalin deceases by 13% and increases by 7%, respectively, from before to after being immersed in formalin. The same trend is found with condition factor in the few papers that report this parameter. Body length decreases on average by c. 2% in fixated and preserved fish regardless of whether the formalin is diluted in seawater or fresh water.     

Effects of freezing on white perch Morone americana (Gmelin, 1789): implications for multivariate morphometrics

This study tested the hypothesis that duration of freezing differentially affects whole‐body morphometrics of a derived teleost. Whole‐body morphometrics are frequently analyzed to test hypotheses of different species, or stocks within a species, of fishes. Specimens used for morphometric analyses are typically fixed or preserved prior to analysis, yet little research has been done on how fixation or preservation methods or duration of preservation of specimens might affect outcomes of multivariate statistical analyses of differences in shape. To determine whether whole‐body morphometrics changed as a result of freezing, 23 whole‐body morphometrics of age‐1 white perch (Morone americana) from western Lake Erie (n = 211) were analyzed immediately after capture, after being held on ice overnight, and after freezing for 100 or 200 days. Discriminant function analysis revealed that all four groups differed significantly from one another (P < 0.0001). The first canonical axis reflected long‐axis morphometrics, where there was a clear pattern of positive translation along this axis with duration of preservation. Re‐classification analysis demonstrated fish were typically assigned to their original preservation class except for fish frozen 100 days, which assigned mostly to frozen 200 days. Morphometric comparisons using frozen fish must be done on fish frozen for identical periods of time to avoid biases related to the length of time they were frozen. Similar experiments should be conducted on other species and also using formalin‐ and alcohol‐preserved specimens.     

Shrinkage of Gobiocypris rarus, Procypris rabaudi, and Sinilabeo rendahli preserved in formalin

Length measurements of preserved fishes are necessary in many types of fish surveys because logistics often do not allow for fish measurement immediately after catch. If the fixative causes significant shrinkage, then the preserved lengths cannot be directly used to indicate accurate live lengths. The objective of this study was to determine how preservation in formalin affects standard length of Gobiocypris rarus larvae (24‐day‐old and newly hatched), larval Procypris rabaudi (4‐day‐old), and larval Sinilabeo rendahli (12‐day‐old). Fishes were measured (to nearest 0.01 mm) and individually fixed in the appropriate formalin solution (2.5% or 5.0% formalin), then re‐measured at 0.5, 1, 3, 7, 14, 30, 45 and 75 days after preservation to follow the time course of shrinkage. Most of the shrinkage occurred within the first half day after preservation. The 5.0% formalin caused a higher relative shrinkage rate than did the 2.5% solution; however, the difference was not statistically significant. In G. rarus, initial shrinkage of newly hatched larvae was higher than that of 24‐day‐old larvae.     

Change in allometric length–weight relationship of Salmo trutta at emergence from the redd

In their life cycle, especially at early life stages, fishes pass through distinct growth stanzas with different length‐weight relationship patterns. Salmonid fish fry emergence from the redd gravel is a crucial moment in their life history. This study presents an ontogenic change in allometric length–weight relationship scaling of sea trout Salmo trutta at the time of emergence from the natural redds in a small lowland stream of western Lithuania. The paper also gives shrinkage rates and correction factors for emergent Salmo trutta fry after protracted formalin preservation.     

Length–length relationship, length–weight relationship, gonadosomatic index, condition factor, size at maturity and fecundity of Parastromateus niger (Carangidae) in Kuwaiti waters

Analyses of morphometric parameters of the black pomfret, Parastromateus niger, in the Kuwaiti waters of the Arabian Gulf during a 44‐month study revealed the following length‐length relationships: TL (cm) = 0.63 + 1.15SL (females), TL (cm) = 0.94 + 1.14SL (males) and TL (cm) = 0.75 + 1.15SL (both sexes including juveniles). The 95% CI of the exponent in length‐weight relationship varied from 2.788 to 2.796 in females, 2.726 to 2.732 in males, 2.779 to 2.784 for both sexes and 2.590 to 2.613 in juveniles, all indicating an allometric relationship. Simultaneous analysis of Fulton’s condition factor (K) and GSI indicate that spawning in black pomfret peaks in May and June even though the fish remain in good condition throughout the spawning season. The Fulton’s condition factor was statistically discriminatory in determining the well‐being of P. niger. Variation in average maximum condition was significant between sexes and also higher in 2004 as compared to 2005. The mean size at first sexual maturity (SL_50%) was 17.5 cm for males and 29 cm for females. Fecundity ranged from 71 305 to 3 895 449 eggs and correlated positively with: (a) ovary‐free body weight (P < 0.05), standard length (P < 0.05) and ovary weight (P < 0.05), and negatively with egg size (P > 0.05). Fecundity‐total length relationship was best described by the logarithmic equation of the form F = 0.898 + 3.15 × TL^3.15. The use of results of such morphometric studies in the management of the P. niger species in the Arabian Gulf is suggested due to its growing commercial importance.     

Effect of starvation and re‐feeding on growth performance and content of plasma lipids,glucose and insulin in cultured juvenile Persian sturgeon (Acipenser persicus Borodin, 1897)

The effect of starvation and subsequent re‐feeding to satiation on compensatory growth performance, insulin and blood serum values were investigated in juvenile Persian sturgeon (Acipencer persicus) with an average weight 108.04 ± 0.28 g (mean ± SEM) and in the same rearing condition over an 8‐week period. Sturgeons were allocated to one of five feeding treatments: controls (C, continuous feeding), W1 (1 week starvation), W2 (2 weeks starvation), W3 (3 weeks starvation) and W4 (4 weeks starvation), followed by a single 4 weeks of re‐feeding to satiation. Changes in growth performance and blood serum indices were examined at the end of weeks 4 and 8. Body weight, specific growth rate (SGR), condition factor (CF) and weight gain were determined to have significantly decreased during starvation. Fish starved for 1 week reached the same weight as the control fish after re‐feeding for 4 weeks, indicating that complete compensatory growth occurred. Although the specific growth rate in W2, W3 and W4 fish was greater than that in the control fish after re‐feeding, W2, W3 and W4 fish did not reach the same body weight as control fish at the end of re‐feeding period, and showed partial compensation only. Blood plasma, glucose and insulin concentrations did not change significantly during starvation and re‐feeding (P > 0.05). This suggests that sturgeon are able to maintain glycaemia during starvation, probably due to their non‐carbohydrate dietary source. Plasma total lipid and triglyceride levels increased in starvation treatments, whereas the increases were significant only in W3 treatment (P < 0.05). After a 4‐week re‐feeding period, their levels decreased in comparison to the starvation periods. Increases in plasma total lipid and triglyceride levels appear to be due to their roles as preferred nutrients for mobilization in Persian sturgeon and the magnitude and duration of compensatory growth depended on the length of food deprivation.     

Effects of sodium chloride on water quality and growth of Oreochromis niloticus in earthen ponds

A 2 × 2 factorial experiment was conducted between April and August 2001 to evaluate the effects of NaCl on Oreochromis niloticus growth and water quality in twelve 0.015 ha limed ponds. The design involved fertilizer and salt as factors with two treatments for each factor. Each salt‐fertilizer combination was replicated three times and fish were not offered external food during the 98 days culture. Growth of O. niloticus was significantly enhanced by salt at higher fertilizer level but not at the lower fertilization level. Although salt had no direct effect on fish growth, a significant salt‐fertilizer interaction was demonstrated. Water quality variables, with a few exceptions, were similar among the salted treatments. Total ammonia increased significantly with fertilization level, but the values were similar in salted and unsalted treatments at the same fertilizer level. Total nitrogen was higher in the salted than unsalted treatments while the organic matter content was lower in the salted treatments. The reasons for the better growth of O. niloticus, are discussed with respect to water quality variables. The present results suggest that fertilization rates of 20 kg N ha^?1 may have negative effects on fish growth. However, presence of sodium chloride seems to reduce these negative effects.     

Shrinkage of Prochilodus lineatus (Valenciennes, 1847) larvae preserved in either ethyl‐alcohol or formalin in relation to their developmental stage and feeding condition

The effects of preservation in 95% ethyl‐alcohol and 5% formalin were analysed for 3 months on standard length of Prochilodus lineatus larvae from hatching to the end of the flexion process. Unyolked stages were raised under two feeding regimes: unfed and daily fed. All developmental stages that were preserved in formalin as well as the yolked and flexion‐postflexion larvae stored in alcohol shrank significantly (2–6%). In contrast, unyolked preflexion larvae showed a slight but significant enlargement after storage in alcohol (1%). Shrinkage of preflexion stages was 2.5% higher when stored in formalin, while both preservative agents caused similar shrinkage in flexion‐postflexion larvae (ca. 3%). Shrinkage levels after storage in alcohol were dependent on live length, decreasing or increasing with increasing length in yolked and flexion‐postflexion larvae, respectively. The feeding regime did not affect length changes after preservation in either preservative agent.     

Studies on the biology of Nile tilapia (Oreochromis niloticus) in Lake Victoria,Kenya: in light of intense fishing pressure

Nile tilapia Oreochromis niloticus is now the most abundant and commercially important tilapiine in Lake Victoria. From the total of 1 512 fish sampled from commercial gill net fisheries during 2014 and 2015, 809 (54%) were males and 672 (44%) were females, giving an overall sex ratio of 1.20 males: 1.00 females. The mean (± SE) length and weight for all fish were 28.7 (±0.1) cm TL and 506.6 (±7.1) g, respectively. The slope b of the length-weight relationship was 2.98, 3.01 3.01, for males, females, and combined sexes, respectively. The relative condition factor was 1.02 for males and 1.04 for females with little variation across the months of sampling. The length at 50% maturity was estimated as 31.0 cm TL for male Nile tilapia and 26.0 cm TL for females. Sixty percent of the fish in the commercial catches surveyed were below 30 cm TL. Comparisons with earlier studies in this system suggest an overall decline in size at maturity over the past 30 years, which may reflect intense fishing pressure.     

Genetic integrity of black sea bream (Acanthopagrus schlegeli) sperm following cryopreservation

Although semen cryopreservation has been applied successfully in many fish species, extensive variation in post‐thaw semen quality exists between species and individuals. AFLP (amplified restriction fragment length polymorphism) is a powerful method for detecting DNA polymorphisms at the individual, population, and species levels. The method has been successfully applied to boars (Sus domestica, Suidae, Artiodactyla, Mammalia) to detect and evaluate differences in DNA sequences that correspond with semen integretiy after employing various freezing techniques. Freezing and thawing of semen has also an effect of selecting for freezing‐resistant (or intact) and eliminating non‐viable or defective sperm. Only the fully intact and functional sperm, despite potential compromise by adverse freezing and osmotic stresses, retain fertility after thawing. The objective of this study was to use AFLP to assess any genetic changes associated with the effect of employed cryo‐methodology on the genetic integrity of sperm of the black sea bream (Acanthopagrus schlegeli) under different cryopreservation treatments. The cryopreservation protocols had no significant effect on sperm motility or survival rate of fertilized ova regardless of using fresh (% motile sperm 89.6 ± 3.0; % embryonic survival rate 54.4 ± 2.9) and frozen‐thawed semen (% motile sperm 80.2 ± 2.0; % embryonic survival rate 51.8 ± 2.0). The post‐thaw sperm motility and survival rates were not significantly different among the sperm samples of the five black sea bream males examined. In the present study, the remaining black sea bream sperm that survive the cryopreservation limit the power of AFLP to trace the genetic markers which correlate with the differences in the sensitivity of sperm to cryo‐injury. It is also possible that point mutations outside the AFLP priming sites may not have been detected. More thorough investigations are needed to determine whether such DNA fingerprints would be found in fish species.     

The growth characteristics and yield potential of rice (Oryza sativa) under non‐flooded irrigation in arid region

The objectives of this field experiment were to study the growth characteristics and yield potential of rice plants under non‐flooded irrigation in arid area. Non‐flooded treatments included drip irrigation with plastic mulching treatments (DIs), furrow irrigation with plastic mulching treatment (FIM) and furrow irrigation with non‐mulching treatment (FIN). Conventional flooded cultivation (F) was check treatment (CK). The four drip irrigation treatments differed in the amount of water applied before and after panicle initiation. Root length density, leaf dry weight, shoot dry weight and root activity were generally higher in the non‐flood‐irrigated treatments (especially the drip‐irrigated treatments) than in the flood‐irrigated treatment at mid‐tillering. However, the growth and development of rice plants were limited after jointing in the non‐flooded irrigation treatments. Increasing the root/shoot ratio and root length density in the 20–40 cm depth and decreasing specific root length at 0–20 cm soil layer were important mechanisms for helping the rice plants to adapt to the non‐flooded environmental stresses. Finally, the grain yield in the non‐flooded irrigation treatments was lower than that in the F treatment. These low yields were mainly attributed to the low root length density at 0–20 cm depth and root activity. Generally speaking, the restricted degrees in the DIs were smaller than that in the FIM and FIN treatments. Among the DIs, both the highest grain yield (8223–8900 kg ha^?1) and the highest water use efficiency (WUE) (0.63) were observed when the soil water content was kept at ?30 kPa before panicle initiation and at ?15 kPa after panicle initiation (referred to as the DI2 treatment). The yield in the DI2 treatment was not significantly different than that in the flood‐irrigated treatment. However, WUE was 2.5 times higher in the DI2 treatment than in the F treatment. These results suggest that drip irrigation technology can be considered as a better water‐saving cultivation of rice plants in arid region.     

The effect of feeding strategies and body weight on growth performance and hematological parameters of Siberian sturgeon (Acipenser baerii,Brandt 1869): Preliminary results

The purpose of present study was to investigate the growth performance and hematological parameters of Siberian sturgeon Acipenser baerii in three feeding strategies (satiation feeding, restricted feeding and starvation). We considered two body weights 465.75 ± 11.18 g and 250.40 ± 12 g for large fish and small fish, respectively. In a 45‐day experiment, eighteen fiberglass tanks were used for six triplicate treatments. In restricted feeding strategy, the fish were fed to 50% of satiation and starved treatments were an absolute feed deprivation throughout the experiment. At the end of 45‐day period, the hematological parameters were measured. The reduction of weight, condition factor and daily growth rate in the smaller fish was dramatically higher than the larger fish. Feed deprivation increased the rate of weight loss. Specific growth rate were negative in the large and small fish of starved treatments. The hematological results revealed no significant differences in except for the number of monocytes in larger fish. Large satiated fish showed the highest number of monocytes. In general, these strategies (restriction and deprivation) did not have any negative impacts on the hematological parameter in both sizes but respect to fish body weight, difference in total feed input between satiated and restricted treatments changed growth performance. It seems that restricted feeding (50% of satiation) in large fish was enough to achieve optimal growth while the amount of feed provided for small fish was not enough. Therefore; we can conclude that the restricted feeding is an effective strategy in rearing sturgeon at higher weight when the rearing condition is unsuitable.     

Length–weight relationship and condition factor of the Characidae matrinxã, Brycon falcatus (Müller & Troschel, 1844), in the Teles Pires River,southern Amazon

The aim of this study was to verify whether the in situ supply of soybean as a fish attractant bears any influence on the length–weight relationship and condition factor of the omnivorous fish species, Brycon falcatus. In addition, estimated was the length–weight relationship, standard and total length, condition factor, and size of B. falcatus in the spawning period in the Teles Pires River Basin. In the experimental design, rivers of the Teles Pires Basin containing soybean attractant (Verde River, Celeste River, Teles Pires River and the Tapaiuna River) were mapped. Fish were also collected from the Cristalino River, which contained no soybean attractant (treatment control). Samples were collected from August 2012 to July 2013. Length–weight relationship of B. falcatus in the Teles Pires River Basin can be obtained by the expression: Wt = 0.0091*SL^3.376, r² = .97, n = 102, and length‐length relationship: TL = 1.128*SL + 3.978, r² = .96. The condition factor of all specimens collected in the Teles Pires Basin was k = 1.055. Size at capable phase (L50) of spawning was estimated at 23 cm standard length. Results showed no significant difference in the length–weight relationship or condition factor of B. falcatus collected from rivers with or without soybean attractant. Concluded is that the in situ supply of soybean as a fish attractant does not interfere with the wellbeing of B. Falcatus in the Teles Pires River Basin.     

The effects of formalin and isopropyl alcohol on length and weight measurements of Sarotherodon mossambicus Trewavas

Two groups of 30 Sarotherodon mossambicus were weighed and measured when alive, after 5 days in 10% formalin, and after 5, 10, 15, 20, 25, 30 and 65 days in 37.5% isopropyl alcohol. Fish from two size classes were preserved and all fish initially increased slightly in length and increased greatly in weight. The greatest increases in length and weight were noted after 5 days in formalin solution. After 65 days in isopropyl alcohol, the length of the preserved fish decreased to approach the live length while the weight of the fish stabilized at a higher than live weight value. This study is the first report of slight length increases or no shrinkage in preserved fish. Comments on the use of standard correction factors for lengthweight studies of preserved fish are provided.     

Reproductive biology, length-weight relationship and relative condition of pond raised Tilapia zilli (Gervais)

In 'runted' populations of Tilapia zilli positive correlations were found between maturation stages and the following: gonadosomatic index, gonad weight, fish weight and fish length. The minimum size at maturity was 9·0 cm in males and 11·0 cm in females. The mean fecundity in the 'stunted' females was 2359 eggs and it increased with length ( L ) weight (HO and depth ( D ) of the fish. There was, however, no correlation between fecundity and egg size. The results indicated that growth in this 'stunted' T. zilli population was not isometric. The relative condition factor ( Kn ) which approached one indicates good condition.     

A new technique for long preservation of 14C-labelled Cladocerans

Three ways of preserving labelled Cladocerans fed with ¹⁴C-Chlorella for 7.5–10 min were tested. Tracer leakage in 4% formalin at room temperature is rapid and extensive (half of the label was found in the animals after 1 hour of preservation). Even when individuals are frozen and sorting is made quickly in a liquid, losses nevertheless occur (substantial decrease of animal activity after only 4–5 min in the water in one of the two experiments performed). Results obtained after freezing in 4% formalin and sorting exactly 2 hours after thawing gave consistent losses: 16 separate experiments with Daphnia pulex, Ceriodaphnia spp. and Diaphanosoma brachyurum gave apparent filtering rates underestimated from 35% to 63% for freezing periods of up to 45 days. The good agreement in in situ community filtering rates between measured values and estimated ones from individual data confirmed the validity of a correction factor of x 2 applied to animals frozen in formalin.     

Length–weight relationships for six fish species in Iskenderun Bay (Eastern Mediterranean Sea coast of Turkey)

The length–weight relationships (LWRs) were estimated for six non‐indigenous fish species, namely, Apogon smithi (Kotthaus, 1970); Ostorhinchus fasciatus (White, 1790); Pomadasys stridens (Forsskål, 1775); Champsodon capensis Regan, 1908; Torquigener flavimaculosus Hardy & Randall, 1983; and Tylerius spinosissimus (Regan, 1908) from Iskenderun Bay, eastern Mediterranean Sea. Their length–weight relationship b values ranged from 2.902 to 3.501, and all regressions were found to be significant for all six species (P < 0.001). This study is the first reference on length–weight relationships for these six non‐indigenous fish species from the Eastern Mediterranean Sea coast of Turkey.     

Condition factor,length–weight and length–length relationships of an endangered fish Ompok pabda (Hamilton 1822) (Siluriformes: Siluridae) from the River Gomti,a tributary of the River Ganga,India

The present study describes the condition factor, length‐weight and length‐length relationships for an endangered freshwater fish species Ompok pabda (Hamilton) of the family Siluridae from the River Gomti in Northern India. The values of regression parameter b ranged from 2.81 to 3.32 (r² > 0.90). During the pre‐monsoon, the allometric coefficient b of the LWR was close to isometric value (b = 3.08) allometric (b = 2.87), although it suggested negative allometric growth in monsoon periods while positive growth in post‐monsoon. The condition factor values ranged from 0.672 to 0.744. Results of the present study could be useful to help in conservation and sustainable fisheries management of this endangered species.