CN111676301B - Method for evaluating temperature and humidity state of environment where nursery pigs are located based on relative abundance of nasal microorganisms - Google Patents
Method for evaluating temperature and humidity state of environment where nursery pigs are located based on relative abundance of nasal microorganisms Download PDFInfo
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Abstract
The invention discloses a method for evaluating the temperature and humidity state of the environment where a nursery pig is positioned based on the relative abundance of microorganisms in nasal cavities.
Description
Technical Field
The invention belongs to the technical field of evaluation of pig breeding environments, and particularly relates to a method for evaluating the temperature and humidity state of an environment where a nursery pig is positioned based on the relative abundance of microorganisms in nasal cavities.
Background
The breeding environment is one of factors determining the breeding productivity of live pigs, the contribution rate to the breeding of the live pigs is up to 25%, and the improvement of the breeding environment is a necessary means for improving the breeding productivity of the live pigs. The temperature and humidity state is the most important part of the cultivation environment, and can be represented by a temperature and humidity index. The warm-wet environment can influence the pig breeding productivity from multiple angles, the immunity of the pigs is reduced under the warm-wet environment of cold stress and heat stress, the fat deposition of the pigs is reduced at high temperature, and the pigs in the non-comfortable environment warm-wet state area need to consume more energy to maintain constant body temperature, so that the feed conversion rate is reduced. The equipment of current regulation plant live pig breeds warm and humid environment has cascade fan cooling system and heat preservation lamp electric fan heater system, can adjust power and the heat-generating heat preservation lamp has been produced, and accurate breed concept also is put forward and accurate environmental control is part in the accurate breed.
The accurate evaluation of the pig raising environment is a prerequisite for precise environment control, and the temperature and humidity environment part of the environment evaluation in the current pig raising process is generally completed by a thermometer and a hygrometer. The worker uses a thermometer and a hygrometer to measure the temperature and the humidity of the pig house, then uses a temperature-humidity index calculation formula to calculate the temperature-humidity index and evaluate the temperature-humidity environment of the pig house by using the temperature-humidity index calculation formula, wherein the temperature-humidity index can be calculated by any two values of dry bulb temperature (Td, DEG C), wet bulb temperature (Tw, DEG C), dew point (Tdp, DEG C) and relative humidity (RH, DEG C), and one of the following calculation formulas is selected: thi=td+0.36tdp+41.2 or thi=0.81 td+ (0.99 Td-14.3) rh+46.3 or thi=0.72 (td+tw) +40.6. The method can judge the temperature and humidity environment state of the breeding house at a certain time point, but the temperature and humidity environment state of the live pigs in a period of time cannot be accurately evaluated. The temperature and humidity states of the breeding house generally change dynamically along with time, and the temperature and humidity states of different areas of the same animal house are different, so that the temperature and humidity indexes measured and calculated by the thermometer and the hygrometer at fixed points at fixed time are difficult to evaluate the temperature and humidity states of live pigs moving in certain areas within a period of time. The growth of the microorganism is influenced by the temperature and humidity state of the environment, and the temperature and humidity state of the environment where the individual animal grows can be reversely deduced from the growth state of the microorganism.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a method for evaluating the temperature and humidity state of the environment where the nursery pig is located based on the relative abundance of nasal microorganisms.
In order to achieve the above purpose, the technical scheme provided by the invention is as follows:
the method for evaluating the environmental temperature and humidity state of the nursery pig based on the relative abundance of the nasal microorganisms comprises the step of evaluating the environmental temperature and humidity state of the nursery pig by simultaneously measuring the relative abundance of microorganisms at the levels of the inner door, class, order, family, genus and species of the nasal cavity of the nursery pig (the relative abundance of microorganisms belonging to the prokaryotic kingdom relative to the total prokaryotic microorganisms, the relative abundance of microorganisms belonging to the eukaryotic kingdom relative to the total eukaryotic microorganisms), wherein the microorganisms comprise: fusobacterium, actinobacillus, leuconostoc, erysipelotomyces, lichtheeiraceae, bergey, klebsiella, megamonas, butyriciococcus, bacteroides, interstinimonas, anaerosporipes, pseudomonas, flavonoides, propionimibacterium, carbon dioxide phaga, chryseolepsis Alkalibacterium, leuconostoc (Leuconostoc), clostridium_disporium species, lactobacillus salivarius (Lactobacillus salis), dorea_longicatena species, ruminococcus_calidus species, eubacterium mucilaginosa (Eubacteim), ochrobactrum_pseudo-strain, alicycobacter_pomomorium species, pseudomonas_indica species, euglena (Euglena), pyruscus, sterilla (Candida albicans), pyruscus_x_bretscheideri species; wherein the bacteria not labeled chinese are because there is no chinese name.
The method comprises the following specific steps:
(1) Establishing and selecting a regression model (selection standard: R) of the relative abundance of nasal microorganisms and the environmental temperature and humidity index of the nursery pig reflecting the temperature and humidity environment state of the nursery pig 2 >0.8, requirement p relating to the p value<0.05 As follows:
THI＝76.44-5.12x 1 -49.90x 2 +183.75x 3 +113467.5x 4 +8.46x 5 +62.63x 6 +167.81x 7 -194.40x 8 -430.14x 9 -845.78x 10 -641.75x 11 +8802.26x 12 +38456.91x 13 +47619.7x 14 -63835.86x 15 +20909.23x 16 +37974.96x 17 +38386.48x 18 -27.50x 19 -1105.19x 20 -1599.53x 21 -3461.54x 22 -1647.85x 23 +451.76x 24 -576.22x 25 +13202.74x 26 -123721.2x 27 -1069.93x 28 -11177.17x 29 -487.09x 30 -129.91x 31 the method comprises the steps of carrying out a first treatment on the surface of the The model is LASSO model, and the coefficient R is determined 2 ＝0.9991；
Wherein THI is the environmental temperature and humidity index, x of the nursery pig 1 To x 31 Represents, respectively, the species Fusobacterium nasalis, actinomyces, erysiphelotrichia, lichtheimaceae, burjie's genus, klebsiella, butyricicoccus, bacteroides, interstinimonas, anaerobiostigs, pseudomonas, flavobacterium, propionimicinum, carbon dioxide-philic bacteriaThe relative abundance of Chryseolinea, alkaliibacterium, leuconostoc, clostridium, lactobacillus salivarius, dorea longicatena, ruminococcus_Callicarus, eubacterium mucilaginosa, ochrobactrum pseudorhizonense, alicmyclobus_pomorum, pseudomonas faciens_indica, ooculopsis, pyrola, sterkella, candida albicans, pyrus x bretscheideri, the relative abundance units are: the%;
(2) Quantitatively detecting the concentration of microorganisms in the nasal cavity of the nursery pig, which is related to the model, and calculating to obtain the environmental temperature and humidity index of the nursery pig in a short period of growth by using a corresponding regression model according to the detected result.
Wherein, the nasal cavity microorganism relative abundance index is obtained by adopting 16S rDNA sequencing (detecting prokaryotic microorganism) and 18S rDNA sequencing (detecting eukaryotic microorganism) of an IonS5TMXL sequencing platform.
The invention is further described below:
the diet and the variety in the breeding process of the nursery pigs are ensured to be consistent, the breath heat measuring cabin is used for ensuring that the pigs are in a stable environment temperature and humidity state and the design is repeated, and the fluctuation of the relative abundance of the nasal cavity microorganisms is only influenced by the environment temperature and humidity state. The invention respectively carries out correlation analysis on the relative abundance index of each nasal cavity microorganism of the nursery pig under different temperature and humidity environment state test conditions and the temperature and humidity index of the corresponding environment, and selects a nasal cavity microorganism single index with stronger correlation with the temperature and humidity environment state to build a model (selection standard: correlation analysis |r|)>0.6，p<0.05 The nasal cavity microorganism relative abundance index is also integrated with the corresponding temperature and humidity index, and a model is established and selected by Matlab software (selection standard: r is R 2 >0.8, requirement p relating to the p value<0.05 The nasal cavity microorganism relative abundance index combination capable of accurately reflecting the temperature and humidity environment state is searched, the temperature and humidity environment state of the nursery pig in a period of time is evaluated by using the nasal cavity microorganism relative abundance index and is represented by a temperature and humidity index predicted value, and the temperature and humidity state evaluation of the growth of the nursery pig is more objective and reasonable.
The animal in the invention is a nursery pig, and can be popularized to other growth stages of live pigs and even other animals and people, the method for detecting the relative abundance of the nasal cavity microorganism is not unique, and other technical means capable of accurately detecting the relative abundance of the nasal cavity microorganism can be replaced.
The single index and index combination of the relative abundance of other nursery pig nasal microorganisms not mentioned in the invention can be used for constructing a model by unitary or multiple regression analysis and environmental temperature and humidity index values; in the production stage of other pigs (fattening pigs, bred pigs and the like), a model can be constructed through single indexes or index combinations of nasal microorganisms of the nursery pigs through unitary or multiple regression analysis and environmental temperature and humidity index values, and the model is used for evaluating the environmental temperature and humidity state; and are also within the scope of the present invention.
Compared with the prior art, the invention has the beneficial effects that:
according to the invention, the temperature and humidity environment (expressed by the temperature and humidity index value) of the nursery pig in a short time in the past can be accurately evaluated by measuring the relative abundance of the corresponding nasal microorganisms of the nursery pig and using the model. The calculated temperature and humidity index of the environment where the nursery pig is located has guiding significance for a manager to adjust factors such as temperature, humidity and the like of the nursery pig house, and the temperature and humidity environment of the nursery pig can be controlled to be always in an optimal state in the production process by combining with the growth environment standard of the nursery pig.
Drawings
Fig. 1 shows the trend of the temperature and humidity index change during the test: the abscissa is time, the ordinate is temperature and humidity index value, the test is started from the first day 12:00 to the next day 6:00, the observation time interval is uniformly 30min (the temperature and humidity environment of the breathing cabin is in a stable state in the observation time period), the temperature and humidity index value in the graph is calculated by dry bulb temperature and relative humidity measured by a temperature sensor and a humidity sensor of the breathing cabin according to the formula THI=0.81 Td+ (0.99 Td-14.3) RH+46.3, and the temperature and humidity index value in a small frame in the graph is the average value in the whole test process (the average value of the temperature and humidity index values observed in the test time period is obtained).
Detailed Description
The experimental methods, materials and reagents used in the examples described below are conventional methods, materials and reagents, unless otherwise specified, and are commercially available.
1. Test animals
40 large-long-white binary hybrid nursery pigs with no significant weight difference are selected and randomly divided into 5 groups, 8 nursery pigs (n=8) in each group are fed into a respiratory cabin in a single column, and enough drinking water is supplied to provide the same reference NRC standard configuration feed for free feeding.
2. Test procedure and sample collection
The temperature and humidity environment of the nursery pigs in the breath heat measuring cabin is controlled, and the temperature and humidity environment of each group is the same, and is controlled by an air conditioner in the test. After the later calculation, the environmental temperature and humidity indexes of the growth of 5 groups of test pigs in the test period are 57.5, 62.1, 74.4, 81.8 and 83.7 respectively (the calculation process is detailed in the description of the drawings). Each test pig was raised in a stable warm and humid environment for about 20 hours, after which the nasal vestibule of the nursery pig was scraped using medical sterilized cotton to collect a nasal microbial sample of the nursery pig.
3. Nasal cavity microorganism sample detection and data analysis
The relative abundance of each level of the prokaryote of the nasal cavity microbial sample was detected using 16S rDNA sequencing of the IonS5TMXL sequencing platform, while the relative abundance of each level of the eukaryote of the nasal cavity microbial sample was detected using 18S rDNA sequencing. Using IBM SPSS Statistics software to make correlation analysis on the relative abundance of single nasal cavity microorganism and test temperature and humidity index, and selecting nasal cavity microorganism single index with strong correlation with temperature and humidity environment state to make model (selection standard: correlation analysis |r|>0.6，p<0.05 Then the nasal cavity microorganism relative abundance index is integrated with the corresponding temperature and humidity index, and a model is established and selected by Matlab software (selection standard: r is R 2 >0.8, requirement p relating to the p value<0.05 Searching for a nasal cavity microorganism index combination capable of accurately reflecting the temperature and humidity environment state.
4. Test results (see Table 1 and FIG. 1)
TABLE 1 environmental temperature and humidity index regression equation for nursery pig by using nursery pig nasal cavity microorganism index
The single index of the relative abundance of the nasal cavity microorganisms cannot establish a regression model which accords with a selection standard with the environmental temperature and humidity index, but it is feasible to evaluate the environmental temperature and humidity index of the nursery pig in a short period by utilizing the combination of the nasal cavity microorganism indexes. The optimal model for evaluating the temperature and humidity state of the environment where the nursery pig grows by utilizing the nasal cavity microorganism combination is as follows: thi=76.44-5.12 x 1 -49.90x 2 +183.75x 3 +113467.5x 4 +8.46x 5 +62.63x 6 +167.81x 7 -194.40x 8 -430.14x 9 -845.78x 10 -641.75x 11 +8802.26x 12 +38456.91x 13 +47619.7x 14 -63835.86x 15 +20909.23x 16 +37974.96x 17 +38386.48x 18 -27.50x 19 -1105.19x 20 -1599.53x 21 -3461.54x 22 -1647.85x 23 +451.76x 24 -576.22x 25 +13202.74x 26 -123721.2x 27 -1069.93x 28 -11177.17x 29 -487.09x 30 -129.91x 31 (comprehensive of levels of phylum, class, order, family, genus, species, LASSO model, determination of coefficient R 2 =0.9925), wherein THI is the environmental temperature and humidity index of the nursery pig, x 1 To x 31 Represents, respectively, the genus Fusobacterium (Fusobacteria), actinomycetes (Actinobacteria), erysipelotomyces, lichtheimaceae, bergey (Bergeella), klebsiella (Klebsiella), megamonas (megamons), butyricicoccus, bacteroides (bacteriodes), interstinimonas, anaerospories, pseudomonas, flavobacterium Propionibacterium, carbon dioxide-bearing fibrinopsis (Capnocytophaga), chryseolinea, alkalibacterium, leuconostoc (Leuconostoc), clostridium_disporococcus, lactobacillus salivarius, dore a_longicatena, ruminococcus_calidus, bacillus mucilaginosus, ochrobactrum_pseudomondsi, alicyclobacillus_pos, pseuudoxantomon as_indica species, euglena (euglena), pyrus, sterkiella, candida albicans (Candid a_albicans), pyrus x bretschneideri species relative abundance (unit:%).
Claims (2)
1. A method for evaluating the temperature and humidity state of the environment where a nursery pig is positioned based on the relative abundance of microorganisms in nasal cavities, which is characterized in that the method evaluates the temperature and humidity state of the growth environment of individual nursery pigs by simultaneously measuring the relative abundance of microorganisms in the inner doors, class, order, family, genus and species levels of the nasal cavities of the nursery pig; the microorganism comprises: fusobacterium gateFusobacteria) The actinomycetes isActinobacteria）、ErysipelotrichiaThe fungus class,LichtheimiaceaeGenus Boehmeria of family JuniperidaeBergeyella) Klebsiella genusKlebsiella) Genus megamonasMegamonas) Butyriciococcus, bacteroides and its preparation methodBacteroides）、IntestinimonasGenus of bacteria,AnaerostipesGenus of bacteria,PseudopedobacterGenus of bacteria,FlavisolibacterGenus of bacteria,PropionimicrobiumGenus of bacteria and carbon dioxide-induced fibrous bacteriaCapnocytophaga）、ChryseolineaGenus of bacteria,AlkalibacteriumGenus LeuconostocLeuconostoc）、Clostridium_disporicumBacterial strain and lactobacillus salivariusLactobacillus_ salivarius）、Dorea_longicatenaBacterial strain,Ruminococcus_callidusBacterial strain, mucus eubacilliEubacterium_limosum）、Ochrobactrum_pseudogrignonenseBacterial strain,Alicyclobacillus_ pomorumBacterial strain,Pseudoxanthomonas_indicaBacterial species, euglena and the likeEuglenida）、PyrusGenus of bacteria,SterkiellaGenus candida albicansCandida_albicans）、Pyrus_x_bretschneideriA strain;
the method comprises the following specific steps:
(1) The regression model of the relative abundance of the nasal microorganisms and the environmental temperature and humidity index of the nursery pig reflecting the temperature and humidity environment state of the nursery pig is established and selected as follows:
THI=76.44-5.12x 1 -49.90x 2 +183.75x 3 +113467.5x 4 +8.46x 5 +62.63x 6 +167.81x 7 -194.40x 8 -430.14x 9 -845.78x 10 -641.75x 11 +8802.26x 12 +38456.91x 13 +47619.7x 14 -63835.86x 15 +20909.23x 16 +37974.96x 17 +38386.48x 18 -27.50x 19 -1105.19x 20 -1599.53x 21 -3461.54x 22 -1647.85x 23 +451.76x 24 -576.22x 25 +13202.74x 26 -123721.2x 27 -1069.93x 28 -11177.17x 29 -487.09x 30 -129.91x 31 the method comprises the steps of carrying out a first treatment on the surface of the The model is LASSO model, and the coefficient R is determined 2 =0.9991；
Wherein THI is the environmental temperature and humidity index of the nursery pig,x 1 To the point ofx 31 Respectively represents the Fusobacterium nasal cavity phylum, actinomycetes, the order of the nursery pig,ErysipelotrichiaThe fungus class,LichtheimiaceaeThe genus Klebsiella, butyriciococcus, bacteroides, bacillus, and Bacillus,IntestinimonasGenus of bacteria,AnaerostipesGenus of bacteria,PseudopedobacterGenus of bacteria,FlavisolibacterGenus of bacteria,PropionimicrobiumGenus bacteria, carbon dioxide-producing bacteria,ChryseolineaGenus of bacteria,AlkalibacteriumGenus Leuconostoc, genus Leuconostoc,Clostridium_disporicumBacterial strain, lactobacillus salivarius,Dorea_longicatenaBacterial strain,Ruminococcus_callidusBacterial strain, eubacterium mucilaginosum,Ochrobactrum_pseudogrignonenseBacterial strain,Alicyclobacillus_pomorumBacterial strain,Pseudoxanthomonas_indicaBacterial species, euglena,PyrusGenus of bacteria,SterkiellaGenus Candida albicans,Pyrus_x_bretschneideriStrain relative abundance in units of: the%;
(2) Quantitatively measuring the relative abundance of the nasal microorganisms of the nursery pigs, taking the measured results into corresponding regression models, and calculating and obtaining the environmental temperature and humidity index of the nursery pigs in a short period.
2. The method of claim 1, wherein the nasal microbial relative abundance index is obtained using 16S rDNA sequencing and 18S rDNA sequencing of an ion S5TMXL sequencing platform.
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