Effect of the climate and some different protein diets on the visitation pattern of flesh and blow flies of

This work compares the effect of the climate and some different protein diets on the activity of flesh and blow fly populations in one of the biggest mountains of Sarah region, south-west Kingdom of Saudi Arabia. Trapping of adult flies revealed the presence of three populations, namely, Engelisca adhamae (Lehrer & AbouZied, 2008) and Liosarcophaga babiyari (Lehrer, 1995), in addition to a single calliphorid species Calliphora vicina (R−D, 1830). Adults of L. babiyari showed a year round activity, with the major peak during August. The species E. adhamae showed its major peak during May and June, while in addition, adults of C. vicina showed spring activity during the period from March to April. Adult activity of L. babiyari showed a direct correlation with both of the temperature and the wind velocity, but it was inversely correlated with the relative humidity. Otherwise, adult C.vicina was directly correlated with rainfall. However, the studied climatic factors showed a very weak effect on the activity of adult E. adhamae. The trophic niches, was predicted to affect male L. babiyari which showed great affinity to be trapped with fish followed by liver then meat. In comparison, male E. adhamae preferred fish, followed by both, liver and beef. Meanwhile, female L.babiyari, showed higher tendency to both, fish and liver compared with both, beef and hash-hash. In contrast, female E.adhamae showed nearly the same tendency to fish, liver and beef. However, male and female C. vicina showed more attraction to liver compared with both fish and beef.

Furthermore, microhabitat selection may be influenced with each species' preference and tolerance of field temperature and humidity regimes (De los Santos et al., 2006).Some species of flies select microhabitats and activity times based on the thermal regimes (Kurahashi, 2001;Henning et al., 2005 andBattan et al., 2007).

Ehab M. AbouZied 134
Meanwhile, other species prefer humid habitats rather than thermal regimes (Davis & Laurence, 1992;Shah &Tanzeela, 2004 andHenning et al., 2005) and several studies reported that some species did not show clear patterns (Cottam et al., 1998 andMulieri et al., 2008).Additionally, the wind may be a new niche variable but Cottam et al., (1998) reported that wind and rain were less important for many species.
It is expected that populations of flesh fly and blow fly species would show particular food preferences in terms of their physiological optima.Hence, some species suffered high mortality rates when fed on liver diet (Charles & Robert, 1988).Other sarcophagid flies preferred fish as breeding substrates, in urban areas, rural areas and forests (Lopes, 1973;D'Almeida, 1986, 1993and D'Almeida & Barbosa, 1996) and other species even preferred faeces or liver.On the other hand, Mulieri et al., (2008) stated that most of the recorded flesh flies showed preference for faeces as bait, in the woodland occurring at the coastline of Buenos Aires, but only in some cases exhibited similar preference for both faeces and liver.Some species preferred liver for breeding in urban areas, but in rural areas preferred fish.These niche changes also, have been reported by other authors working in, different geographical areas (D'Almeida & Barbosa, 1996;Thomas, 1951 andNorris, 1965).Yin et al., (1994) and(1999), reported on the need for liver as a diet, which was essential for the activation of the sexual receptivity and supported full oocyte development.
Despite the importance of these flies, the fauna of the adult carrion flies of Al-Baha Province (Kingdom of Saudi Arabia) was not studied before.Therefore, the objective of the present work is, to study the effect of the macroclimate and some different protein diets on the activity of the flesh and the blow fly populations inhabiting this area.These factors were predicted to affect the seasonal patterns of adult flies populations.The author examined the hypothesis of the niche segregation (Pianka & Huey, 1978;Kuusela, 1983 andRohde, 2005).Such hypothesis was explored in relation to the environmental factors, for which, analyses of the seasonal activity patterns and its' relationship with climatic parameters, were carried out.At last, the author explored the hypothesis that the coexistence of species, also, required trophic niche segregation, for which the activity of the fly populations, inhabiting this area in relation to their attraction to the baited traps, was studied.

The study area
Al-Baha Province (11,000 square km) is situated in the south west of the Kingdom of Saudi Arabia between Mecca and Aseer (Fig. 1).Although this province is divided into three distinct regions, collection was focused in the region, which is called Sarah.It is characterized by high mountains with temperate weather and rich plant cover resulting from a relatively high annual rainfall.The second region, Tohama, is situated west of Sarah.Tohama is a low land area, which is characterized by very hot humid weather with very little rainfall season.The third region, the eastern hills, 1550-1900 m asl, which is characterized by cool winter, hot summer and poor plant cover.

Climate of Al-Baha
Climatic conditions (mean monthly values for the study period), such as the temperature, rainfall, wind velocity and relative humidity, were all obtained from Al-Baha station of weather and climate (20º17′4′′N, 41º38′35′′E; 1652 m asl).According to the metrological data, the climate of Al-Baha showed a characteristic pattern (Fig. 2).The rainfall was recorded all the year round with three peaks.The most high amount of rainfall was recorded during spring months (March, April and May), with the peak of maximum amount of rainfall, during April (42.9 ml/m 2 ).During winter, especially January, the peak of rainfall amount was 20.6 ml/m 2 .During autumn months, the maximum amount of rainfall was recorded, during October (13.1 ml/m 2 ).The rainfall amount during the summer season was nearly the same during both July and August (10.2and 9.8 ml/m 2 ), respectively.
The temperature scale, showed a very narrow range of fluctuations with a high range of temperature (29 o C − 27.6 o C), during the period from June to September.The moderate temperature scale was recorded, during spring months (20 o C − 26 o C).The threshold of the warm season (26 o C), was recorded during May and the threshold of low temperature scales (15.8 o C), was recorded during January (Fig. 2).
The relative humidity curve showed an inverted elbow shape.The higher rates of relative humidity were achieved during January (55%), December (53%) followed by November (49%).The spring season showed a moderate humidity values (42 − 47 %).However, the least values of relative humidities were recorded during June and September (23%), followed by July (25%).
The scale of wind velocity seemed to be linear or regular all the year round, except from May to September.During this period, there was a slight increase in the values ranging between 8, 9.75 and 8.8 km/h, during June, July and August, respectively.Meanwhile, the least values of the wind velocity were recorded during November, December and January being 5.13, 5.63 and 6.4, km/h, respectively.

Collection of the adult flies
The adult flies were collected using baited traps placed at Gebel Al-Baher (Sarah region).The site of trapping was determined by GPS (20º0′50.4′′N,41º27′43.56′′E;2209 m asl).The traps were made of, a top, inverted transparent plastic container (500 ml), with a yellow cap like bottom, its base possess a twisted spiral valve to close the top and the bottom firmly.In between the top and the bottom, a yellow plastic cone is found with a narrow hole, near the top of the container.Four traps were used during this study.Traps were placed in, one row, 10 meters apart and hanged at, 1.5 meters height from the soil.Three traps, out of the four used, traps were provided with fresh beef, whole fish and liver.The fourth trap was provided with a commercial product named "Hash-Hash", simulating the odor of decaying organic matter.
Traps were transported to the insectery house of the essential sciences department, Community College, twice weekly.After freezing of the collected alive flies for 30 minutes, the traps were emptied.Catch was recorded, taking into consideration, the sex and the species.Finally, baits had been renewed and the traps were returned back again to the site of the collection at Gebel Al-Baher.Collection of the flies started during January ( 2008) and continued for one year round, finished in December, 2008.For accuracy and more confirmation of the data, collection was repeated for a second year round, from January-December, 2009.Data were treated, statistically, as one unit, the average of the two years, especially for the linear regression analysis.The average of the monthly catch per trap was calculated, for the two years (n = 12).Therefore, the curve of the adult abundance was estimated, determining the period of maximum abundance of each species.

Attractivity of the adult flies to different baits
During the month of the maximum abundance of each species, Second year round 2009, baited traps were rotated in position and arrangement, day after day.The catches were recorded daily (n = 30), taking into consideration, number species, sex and type of the bait.

Statistical analyses
Analyses of the data were carried out using "SPSS" version (11).Either (ANOVA) test or, simple linear regression test (Person's correlation test), was used for the suitable estimation of the data obtained.

Activity of the adult carrion flies populations
From the results obtained (Table 1 and Fig. 2), it appeared that, L. babiyari population activity was affected by temperature, relative humidity and wind velocity.During February-March (Fig. 3) adult flies appeared in minor catches (Seven flies/trap/month).As the temperature rose during April and June, a steady increase in the mean catch of the adult flies, was observed.The adult catches were, 38 and 64 flies/trap/month, respectively.The highest peak of L. babiyari flies was recorded during August (at 29.2 o C, 28% and 8.8 knots/hour) reaching 142 flies/trap/month.During September (27.6 o C), a decline in the activity of L. babiyari population was recorded (99 flies/trap/month), followed by a sharp and sudden decrease, during October (31 o C), as only nine flies/trap/month.During November and December (19.3 o C and 17.2 o C), an average of one fly/trap/month, was recorded.In January, adults of L. babiyari disappeared completely from Gebel Al-Baher.Pearson's correlation test revealed a positive significant correlation (r = 0.843, P < 0.01) between the adult activity of L. babiyari and the temperature scale.Both of the relative humidity and the wind velocity significantly affected the activity of L. babiyari adults showing a negative correlation with the relative humidity scale (r = − 0.758, P < 0.01) and a positive correlation (r = 0.67, P < 0.05), with wind velocity, (Table1).In case of E. adhamae and C. vicina (Fig. 3), the activity of adult population showed a very short period during the year round.The activity period of E. adhamae adults started with the emergence of adults during spring months (20−29 o C) March, April and May, reaching the maximum adult activity during June (41 flies/trap/month).The adult activity was reduced to its minimum values, during the hot months of July, August and September (27 o C− 29 o C) (Fig 2).The temperature, rainfall, and wind velocity insignificantly (P > 0.05) affected the activity of E. adhamae adults showing a positive correlation of 0.503, 0.257 and 0.180, respectively.Although, a negative correlation (−0.438) was observed with E. adhamae adults, relative humidity, also, insignificantly affected the adult activity (Table 1).
In comparison, during February, the activity of C. vicina adults started in low numbers (Eight flies/trap/month), then increased in number during March (32 flies/trap/month), reaching the maximum during April (37 flies/trap/month).The adult population of C.vicina started to decrease from May (11 flies/trap/month) to July (one fly/trap/month).The adults completely disappeared during the period from August up to January (Fig. 3).  1) and figure (3) showed that, rainfall was highly affecting the activity of adult C. vicina (r = 0.731 and P < 0.01).However, both of the wind velocity and the temperature showed negative and least insignificant effect on C. vicina adults (r = − 0.054 and r = -0.121,respectively).Meanwhile, relative humidity showed insignificant but, positive correlation (r = 0.22) with the activity of adult C. vicina.

The attraction of adult fly populations to different baited traps
Data represented in table (2) revealed that, the fish baited trap was the foremost, first rank, attractant to L. babiyari adults (8.67 flies/trap/day).Fish baited traps showed significant difference (P < 0.05), when compared with liver (5.73 flies/trap/day), beef (3.6 flies/trap/day) as well as Hash-Hash (0.73 flies/trap/day).Adult flies of L. babiyari showed higher attraction (the second rank) towards liver when compared with the catch of the beef baited trap.A significant difference (P < 0.05) was recorded between the catches of the liver and the beef baited traps.The same conclusion was true for L. babiyari males, as fish attracted more significant (p < 0.05), number of males (3.53 males/trap/day) compared with liver (1.87 males/trap/day), beef (1.4 males/trap/day) and Hash-Hash (0.13 males/trap/day).However, both the fish and the liver baited traps showed nearly, the same attraction to female L.babiyari since, insignificant difference (P > 0.05) was recorded between the catch of the two former traps (4.93 and 4.53 female/trap/day, for fish and liver, respectively).Both of the beef and Has-Hash baited traps showed the least level of attraction (2.2 and 0.6 female/trap/day) to females of L. babiyari but with significant preference (P < 0.05) to beef compared with Hash-Hash.
The adult population of E. adhamae showed no preference to offered baits, with little insignificant tendency towards fish (2.87 lies/trap/day) followed by liver (2.6 flies/trap/day) then beef (2 flies/trap/day).The same conclusion was true in case of E. adhamae females.The average daily catch's was insignificantly (P> 0.05) different among fish (1.57female/trap/day), liver (1.97 female/trap/day) and beef (1.80 female/trap/day).However, E. adhamae males showed significant (p < 0.05) tendency for fish (1.25 male/trap/day), compared with, liver (0.70 male/trap/day) and beef (0.20 male/trap/day).Both, the liver and beef baited traps showed nearly the same attraction (the second rank) for E. adhamae males (P > 0.05).Hash-Hash showed the least significant attraction to E. adhamae males and females.
The calliphorid C. vicina adult population showed a significant tendency (p < 0.05) towards liver (4.07adults/trap/day) when compared with both catches of fish (1.13 adults/trap/day) and beef (0.27 adults/trap/day).Males of C. vicina showed Effect of the climate and some different protein diets on the visitation pattern 139 139 significant tendency (P < 0.05) to liver (1.6males/trap/day), when compared with both fish (0.73 males/trap/day) and beef (0.07 males/trap/day).Also, female C. vicina preferred liver (2.57females/trap/day) more significantly (P < 0.05), than both, fish (0.6 females/trap/day) and also beef (0.2 females/trap/day).Both sexes of C. vicina, showed no catches in traps baited with Hash-Hash during the study period.The Hash-Hash baited trap was the least significant attractant to the three adult fly populations (Table 2).It is not recommended to use this product to attract adult flesh and blow flies.

DISCUSSION
It could be assumed that these three species did not show conservatism of ecological niches in evolutionary time, i.e. phylogenetic history, (Peterson et al., 1999 andDe los Santos et al. 2006) since, different selective pressures have probably led to the development of different life cycles (behaviour, diapause, egg maturation, etc.), at environmental fluctuations.
The study showed that these three species, which are not congeneric, have very different life histories that likely evolved in response to conservatism of thermal optima, across a highly variable temperature environment.Hence L. babiyari disappeared completely from Gebel Al-Baher area during January, meanwhile E. adhamae adults disappeared during the period from September up to February, and C. vicina adults were not present from August up to January.Similarly, Tachibana and Numata (2006) mentioned that the flesh flies were not captured during winter as flesh flies have a pupal diapause.Also, Kurahashi and Ohtaki (1989) and Kurahashi (1997b) reported that Sarcophaga sp. also disappeared in winter.However, Mulieri et al., (2008) stated that the two flesh flies Oxysarcodexia paulistanensis (Mattos, 1919) and Oxysarcodexia varia (Walker, 1836), showed their highest abundance in December and February.Microcerella muehni (Blanchard, 1939), displayed a different seasonal pattern with the abundance in September and October.In comparison, Gabre and AbouZied (2003) collected C. vicina only during the spring season, in both urban and rural areas of Suez province, Egypt.Henning et al., (2005) mentioned that C. vicina was abundant in March-May during two years survey in North Island, New Zealand.Tachibana and Numata (2006) captured a closely related species, Calliphora nigribarbis (Vollenhoven, 1863), between October and mid June, but not in summer season.Meanwhile, Chrysomya megacephala (Fabricius, 1794) population showed the highest activity during the period from September to October.
The authors also, reported seven species of calliphorid flies showing the maximum abundance during both spring and autumn.Our results showed the presence of one activity peak for C. vicina at Al-Baha (20º17′4′′N) suggesting that this species can not over winter in regions at or above 30 o N, because of the low temperature, which restricted their growth (Kurahashi, 2001).The presence of this species in such regions is caused by temporal invasions, from late summer to autumn (Kato, 1960;Miyazaki, 1960 andKurahashi, 2001).Similarly, C. megacephala population showed a single peak per year (Tachibana and Numata, 2006), however seven species of calliphorid flies showing bimodal pattern of abundance during both spring and autumn.
The temperature as well as the relative humidity scale showed a positive and a negative, significant correlation with the activity of adult L. babiyari.Similarly, the abundance of other sarcophagid flies (O. varia, Tricharaea (Sarcophagula) occidua (Fabricius, 1794) and O. paulistanensis), was influenced by minimum and maximum daily temperature, as well as, the night humidity (Henning et al., 2005).
Meanwhile, in case of E. adhamae, the relative humidity scale showed a negative insignificant correlation.Similarly, Shah and Tanzeela (2004) showed no specific visitation pattern for Sarcophaga sp. with respect to changes in the relative humidity.
Also, Mulieri et al., (2008) confirmed that O. varia did not show a clear pattern towards the relative humidity scale.
The hypothesis of the conservative evolution in ecological niches, (Peterson et al., 1999 andDe los Santos et al., 2006), may be applied to these three species.This would explain the similarity between the thermal and trophic preferences found in this study, compared with the range of values reported by other authors.Thus, in relation to the functional component of the niche, results showed that, adult population of L. babiyari preferred whole fish as a bait, when compared with both liver and beef.
Meanwhile, adult population of E. adhamae showed nearly the same tendency to fish, liver and beef.However, female L. babiyari showed nearly the same tendency to both fish and liver followed by beef.In contrast, female E. adhamae, showed nearly the same tendency towards fish, liver and beef.Lopes (1973) and D'Almeida (1986& 1993) reported that Peckia chrysostoma (Wiedemann 1830) preferred fish as breeding substrates, in urban areas, rural areas and forests.Also, D' Almeida, (1993) stated that Sarcodexia innota (Walker, 1861) preferred fish as a substrate more than liver.
Meanwhile, Sarcophagula sp.preferred faeces as collection substrate.D'Almeida and Barbosa (1996) reported that, mashed fish was a highly attractant diet to the larvae of the sarcophagid fly Ravinia belforti (Prado & Fonseca, 1932).
On the other hand, males of L. babiyari showed a great Preference to fish followed by liver then beef.In comparison, male E. adhamae preferred fish as first rank followed by liver and beef as the second rank.
Since E. adhamae (Lehrer and AbouZied, 2008), is recently described species from Gebel Al-Baher, no data are available about its food preference.Further detailed investigations and studies are required, especially, for the species E. adhamae and also, for L. babiyari, to determine which protein material is essential for the males and the females of each population.
Data also revealed that, adult C. vicina showed great attractivity towards liver as a bait when compared with both fish and beef.Similar results were obtained by D' Almeida (1986Almeida ( , 1993) ) for C. megacephala, Fannia sp. and Phaenicia eximia (Wiedemann, 1819), as both preferred bovine liver, followed by fish as a breeding media.Also, Synthesiomyia nudiseta (Van der Wulp, 1883) preferred liver for breeding in urban areas, but in rural areas S. nudiseta preferred fish.However, mashed fish was a highly attractant diet to the larvae of C. megacephala (D'Almeida and Barbosa, 1996).However, both of Thomas (1951) and Norris (1965) found that C. megacephala preferred faeces.
Effect of the climate and some different protein diets on the visitation pattern 141 141 Data also revealed that both males and females of C. vicina showed higher tendency to liver, followed by fish and beef.Yin et al., (1994) and(1999) reported that, more than 10 mg of dietary liver was required for each Phormia regina (Meigen,1826) male to result in 80% insemination of the female and 20 mg of liver was required for each P. regina female, to allow 78% of females to become inseminated.Between 10-15 mg of protein meal (liver), was required to activate the sexual receptivity in 71% of the P. regina females, while between 15-20 mg of liver was needed to support full ooyte development in 70% of the females.
As a final conclusion, males of either calliphorid, or sarcophagid flies, were attracted to the expected niches (baited traps, in this study), where the females are available, either for breeding, feeding, or deposition of eggs.
In summary, this work had shown some segregation of niches between species of flies, based on weather conditions and food availability, but with a high degree of overlap.The preferences found in this area were very similar to those described for these groups of flies in other geographic regions, supporting the hypothesis of conservation of ecological niches in the colonization of the mountainous habitats, facilitating the coexistence of a definite species.

Fig. 1 :
Fig. 1: Location of study site in Gebel Al-Baher, Al-Baha Province, Kingdom of Saudi Arabia.The numbers in abscissas (latitude as the angular distance, in degrees of a point north of the Equator) and ordinates (longitude as the angular distance, in degrees, of a point east of the Prime -Greenwich-Meridian) are used to identify locations on the Earth.
climate and some different protein diets on the visitation pattern 137 137

Table ( 1
): The effect of the climate on the activity of adult carrion flies populations inhabiting Gebel Al-Baher, K.S.A Fig.2: Climate of Al-Baha Province during the study period.Seasonal distribution in monthly average of mean temperature in ºC (white squares), rainfall in ml/m 2 (black squares), relative humidity in % (white circles) and wind velocity in km/h (black circles).

Table ( 2
): Attractivity of baited traps to adult carrion fly populations inhabiting Gebel Al-Baher, Al-Baha Province, K.S.A Mean = catch/ trap/day, (s.e) = the standard error.Symbols in each row, which are different, refer to significant difference (P<0.05) between the values of each baited trap mean, examined by Post Hoc test.No comparisons were constructed between columns, different sexes and species.