Protection of Animals Used for Scientific Purposes

L.N. 297 of 2025

ANIMAL WELFARE ACT

(CAP. 439)

Protection of Animals Used for Scientific Purposes

(Amendment No. 2) Regulations, 2025

IN EXERCISE of the powers conferred by article 7(2) of the

Animal Welfare Act, the Minister responsible for veterinary services,

has made the following regulations:-

(1) The title of these regulations is the Protection of

Animals Used for Scientific Purposes (Amendment No. 2)

Regulations, 2025 and these regulations shall be read and construed as

one with the Protection of Animals Used for Scientific Purposes

Regulations, hereinafter referred to as the "principal regulations".

(2) The scope of these regulations is to transpose Commission

Delegated Directive (EU) 2024/1262 of 13 March 2024 amending

Directive 2010/63/EU of the European Parliament and of the Council

as regards the requirements for establishments and for the care and

accommodation of animals, and as regards the methods of killing

animals.

Annex III to the principal regulations shall be amended as

follows

(a)

Section A thereof shall be amended as follows

(i) the heading of sub-item 2.3. thereof shall be

substituted by the following new item

"2.3. Noise and vibration";

(ii) immediately after paragraph (c) of sub-item 2.3.

thereof there shall be added the following new paragraph

"(d) For aquatic animals, equipment causing

noise or vibration, such as power generators or

filtration systems shall not adversely affect animal

welfare.";

(iii) the heading of sub-item 2.4. thereof shall be

substituted by the following new heading

"2.4. Alarm systems and contingency plans";

(iv) immediately after paragraph (c) of sub-item 2.4.

thereof there shall be added the following new paragraph

"(d) Effective contingency plans shall be in

place to ensure the health and welfare of the animals

in the case of a failure of essential husbandry

systems.";

(b)

Section B thereof shall be amended as follows

(i)

immediately after the first paragraph of item 8

thereof there shall be added the following paragraph

"When housing birds taken from the wild, the

space allowance provided for in tables 8.1 to 8.10

shall apply whenever the birds are held for periods of

more than 24 hours. When birds are held for shorter

periods of time, appropriate measures shall be taken

to minimise, as far as possible, any risks to animal

welfare.";

(ii) immediately after table 8.7 of item 8 thereof

there shall be added the following new tables

"Table 8.8.

Starlings

Minimum

Minimum

Minimum

Minimum

enclosure

height

length of food

length of perch

size

trough per bird

per bird

(cm)

(m 2 )

(cm)

(cm)

2,0

4,0

6,0

0.25

0,15

Table 8.9.

House sparrows

Group size in the

Group size in the

Minimum

Minimum

absence of visual

presence of visual

enclosure size

height

barriers

(m 2 )

(cm)

up to 15

2,4

16 to 35

4,8

36 to 60

7,3

for each additional

for each additional

0,11

bird above 30 in

bird above 60 in

number

Table 8.10.

Great tit and Blue tit

Minimum

Minimum

Minimum

Minimum

enclosure size

height

number of

length of perch

per bird

feeders

per bird

(cm)

(m 2 )

(cm)

100 ";

(iii) sub-item 11.1 thereof shall be substituted by the

following sub-item

"11.1. Water supply and quality

Adequate water supply of suitable quality shall

be provided at all times. Water flow in re-circulatory

systems or filtration within tanks shall be sufficient to

ensure that water quality parameters are maintained

within acceptable levels, according to the

characteristics of the husbandry system, and

according to the requirements of the species and their

life stage requirements. Water supply shall be filtered

or treated to remove substances harmful to fish,

where necessary. Water-quality parameters shall at all

times be within the acceptable range that sustains

normal activity and physiology for a given species

and stage of development. The water flow shall be

appropriate to enable fish to swim naturally and to

maintain normal behaviour. Fish shall be given an

appropriate time for acclimatisation and adaptation to

changes in water-quality conditions. Appropriate

measures shall be taken to minimise sudden changes

in the different parameters affecting water quality.

Appropriate water flow and the water level shall be

ensured and monitored.";

(iv) sub-item 11.2 thereof shall be substituted by the

following new sub-item

"11.2. Oxygen, nitrogen compounds, carbon

dioxide, pH, and salinity

Oxygen concentration shall be appropriate to

the species and to the context in which the fish are

held. Where necessary, supplementary aeration of

tank water shall be provided, depending on the

husbandry system. The concentrations of carbon

dioxide and of nitrogen compounds, namely

ammonia, nitrite and nitrate, shall be kept below

harmful levels. Water quality shall be monitored

using a defined testing schedule at a sufficient

frequency to detect changes in these critical

parameters and action shall be taken to mitigate such

changes.

The pH level shall be adapted to the species and

monitored to be kept as stable as possible. The

salinity shall be adapted to the requirements of the

fish species and to the life stage of the fish. Changes

in salinity shall take place gradually.";

(v) sub-item 11.3 thereof shall be substituted by the

following sub-item

"11.3. Temperature and lighting

Temperature shall be maintained within the

optimal range for the fish species and their stages of

development and kept as stable as possible. Changes

in temperature shall take place gradually. Fish shall

be maintained on an appropriate photoperiod.";

(vi) sub-item 11.5 thereof shall be substituted by the

following new sub-item

"11.5. Feeding and handling

Fish shall be fed a diet suitable for the fish at an

appropriate feeding rate and frequency. Particular

attention shall be given to feeding of larval fish

during any transition from live to artificial diets. If

feed withdrawal is necessary for non-procedural

reasons (e.g. transport), the duration shall be kept as

short as possible and take into account the fish size

and water temperature.

Where possible, fish shall be handled without

removal from the water. Handling of fish both in and

outside of the water shall be kept to a minimum and

equipment in direct contact with the fish shall be

moistened. Fish shall not be handled at the outer

limits of the water temperature ranges that they may

tolerate.";

(vii) immediately after sub-item 11.5 thereof, as

substituted, there shall be added the following item

"11.6. Zebrafish

1.6.1. Water Quality

Table 11.1

Water parameter requirements in zebrafish housing systems

Minimum-maximum requirements

Temperature

24-29°C

150-1700 μS/cm 2

Conductivity

Total Alkanility

40-250 mg/L CaCO 3

6,5-8

NH 3 /NH 4 +< 0,1 (*2) mg/L, NO 2 - < 0,3 mg/L,

NO 3 - < 25 mg/L

> 5 mg/L

10 għandhom japplikaw kull meta l-għasafar

jinżammu għal perjodi ta’ aktar minn 24 siegħa. Meta

l-għasafar jinżammu għal perjodi iqsar, għandhom

jittieħdu miżuri xierqa sabiex jitnaqqsu kemm jista’

jkun possibbli, ir-riskji għat-trattament xieraq tal-

annimali.";

(ii) minnufih wara t-tabella 8.7 tal-partita 8 tiegħu

għandhom jiġu miżjuda t-tabelli ġodda li ġejjin

"Tabella 8.8.

Sturnell

Daqs

Għoli

Tul minimu tal-

Tul minimu tal-

minimu tal-

minimu

ħawt tal-għalf

perċa għal kull

kumpartim-

għal kull għasfur

għasfur

(cm)

ent

(cm)

(cm)

(m )

2,0

4,0

6,0

għal kull

0.25

għasfur

addizzjonali

bejn 21 u 50

għal kull

0,15

għasfur

addizzjonali

meta l-

għadd

jaqbeż 50

Tabella 8.9.

Għasafar tal-Bejt

Daqs tal-grupp

Daqt tal-grupp fil-

Daqs minimu tal-

Għoli minimu

fin-nuqqas ta’

preżenza ta’

kumpartiment

(cm)

ostakli viżivi

ostakli viżwali

(m 2 )

sa 10

sa 15

2,4

11 sa 20

16 sa 35

4,8

21 sa 30

36 sa 60

7,3

0,11

għal kull għasfur

għal kull għasfur

addizzjonali meta

addizzjonali meta

l-għadd jaqbeż 30

l-għadd jaqbeż 60

Tabella 8.10.

Fjorentin u Primavera

Daqt tal-

Daqs minimu

Għoli

Għadd

Tul minimu

grupp

tal-

minimu

minimu ta’

tal-perċa għal

kumpartiment

ħwat tal-

kull għasfur

(cm)

għal kull

għalf

(cm)

għasfur

(m 2 )

2-10

(tal-

istess sess)

100 ";

1 mara + 1

raġel

(iii) is-subpartita 11.1 tiegħu għandha tiġi sostitwita

bis-subpartita ġdida li ġejja

"11.1. Provvista u kwalità tal-ilma

F’kull ħin għandu jkun hemm provvista

adegwata ta’ ilma ta’ kwalità adatta. Il-fluss tal-ilma

f’sistemi ta’ riċirkulazzjoni jew filtrazzjoni f’tankijiet

għandu jkun biżżejjed li jiżgura li l-parametri tal-

kwalità tal-ilma jinżammu f’livelli aċċettabbli, skont

il-karatteristiċi tas-sistema ta’ trobbija u skont ir-

rekwiżiti tal-ispeċi u tal-istadju tal-ħajja tagħhom.

Fejn ikun meħtieġ, il-provvista tal-ilma għandha tiġi

ffiltrata jew ittrattata biex jitneħħew is-sustanzi li

jagħmlu l-ħsara lill-ħut. Il-parametri tal-kwalità tal-

ilma għandhom f’kull ħin jinżammu fil-firxa

aċċettabbli li ssostni l-attività normali u l-fiżjoloġija

tal-ispeċi u l-istadju tal-iżvilupp partikolari. Il-fluss

tal-ilma għandu jkun xieraq li jippermetti lill-ħut

jgħumu b’mod naturali u jġibu rwieħhom b’mod

normali. Il-ħut għandu jingħata biżżejjed ħin sabiex

jikklimatizza u jadatta għal tibdil fil-kondizzjonijiet

tal-kwalità tal-ilma. Għandhom jittieħdu miżuri

xierqa sabiex jiġu mminimizzati l-bidliet f’daqqa fil-

parametri diversi li jaffettwaw il-kwalità tal-ilma. Il-

fluss xieraq tal-ilma u l-livell xieraq tal-ilma

għandhom jiġu żgurati u mmonitorjati.";

(iv) is-subpartita 11.2 tiegħu għandha tiġi sostitwita

bis-subpartita ġdida li ġejja

"11.2. Ossiġnu, komposti

tan-nitroġenu,

diossidu tal-karbonju, pH u salinità

Il-konċentrazzjoni tal-ossiġenu għandha tkun

adatta għall-ispeċi u l-kuntest li fihom jinżamm il-

ħut. Fejn ikun meħtieġ, għandha tiġi pprovduta

arjazzjoni supplimentari lill-ilma tat-tank, skont is-

sistema tat-trobbija. Il-konċentrazzjonijiet tad-

diossidu tal-karbonju u tal-komposti tan-nitroġenu,

jiġifieri l-ammonijaka, in-nitrit u n-nitrat, għandhom

jinżammu taħt il-livelli ta’ ħsara. Il-kwalità tal-ilma

għandha tiġi mmonitorjata bl-użu ta’ skeda tal-

ittestjar definita bi frekwenza suffiċjenti sabiex jiġu

identifikati bidliet f’dawn il-parametri kritiċi u

għandha tittieħed azzjoni sabiex jittaffew tali bidliet.

Il-livell tal-pH għandu jiġi adattat għall-ispeċi u

mmonitorjat sabiex jinżamm kemm jista’ jkun

stabbli. Is-salinità għandha tiġi adattata għall-ħtiġijiet

tal-ispeċi tal-ħut u għall-istadju tal-ħajja tal-ħut.

Tibdil fis-salinità għandu jsir gradwalment.";

(v) is-subpartita 11.3 tiegħu għandha tiġi sostitwita

bis-subpartita ġdida li ġejja

"11.3. Temperatura u dwal

It-temperatura għandha tinżamm fil-limiti

ottimali għall-ispeċi tal-ħut u l-istadji tal-iżvilupp

tagħhom u tinżamm kemm jista’ jkun stabbli. Tibdil

fit-temperatura għandu jsir gradwalment. Il-ħut

għandu jingħata perjodi ta’ dawl adatti.";

(vi) is-subpartita 11.5 tiegħu għandha tiġi sostitwita

bis-subpartita ġdida li ġejja

"11.5. Għalf u immaniġġjar

Il-ħut għandu jingħalef b’dieta li tgħodd għalih

u b’rata u frekwenza adatta. Għandha tingħata

attenzjoni partikolari għall-għalf tal-ħut larvali waqt

kwalunkwe tranżizzjoni minn dieta ħajja għal waħda

artifiċjali. Jekk ikun meħtieġ li jitneħħa l-għalf għal

raġunijiet mhux proċedurali (eż. it-trasport), it-tul ta’

żmien għandu jkun qasir kemm jista’ jkun u jqis id-

daqs tal-ħut u t-temperatura tal-ilma.

Fejn possibbli, il-ħut għandu jiġi mmaniġġjat

mingħajr ma jitneħħa mill-ilma. L-immaniġġjar tal-

ħut kemm fl-ilma kif ukoll barra mill-ilma għandu

jinżamm għall-minimu u t-tagħmir f’kuntatt dirett

mal-ħut għandu jiġi mxarrab. Il-ħut ma għandux jiġi

mmaniġjat fil-limiti lil hinn mill-meded tat-

temperatura tal-ilma li jistgħu jittolleraw.";

(vii) minnufih wara s-subpartita 11.5 tiegħu, kif

sostitwita, għandha tiġi miżjuda l-partita ġdida li ġejja

"11.6 Għasafar taż-żebra

6.2. Lighting

During the light phase, the light levels shall be constant, except

during short dawn and dusk transitions. Dark phase shall be completely

11.6.3. Stocking density and environmental complexity

Water volumes smaller than 1 litre shall not be used for adult

zebrafish. Stocking densities shall not exceed 10 adult fish for every

litre. The tank size and shape shall allow the fish to express their natural

behaviours and swimming activities.

Prolonged single housing shall be avoided.

*2 or below detection limit. 0,1 mg/L indicates the total amount of ammonia, NH /

NH 4 + . This corresponds to 0,002 mg/L of NH 3 at 28 °C and pH 7,5. ";

(viii) immediately after sub-item 11.6.3 thereof there

shall be added the following new item

"12. Cephalopods

1. Water supply and quality

Adequate water supply of suitable quality shall be provided at all

times.

The tank design and water flow rate shall meet the needs of the

animal including appropriate oxygenation delivered in relation to its

size, life stage and behavioural needs. Water temperature, salinity, pH,

and nitrogen compound levels shall be appropriate to the needs of

species and life forms. Escapes and the inadvertent introduction of

foreign elements shall be prevented through the use of covers where

necessary.

Cephalopods shall be given appropriate time for acclimatisation

and adaptation to changes in water quality conditions.

12.2. Lighting

Light intensity and photoperiod shall satisfy the requirements of

the species.

12.3. Diet

Cephalopods shall be provided with a feeding regime appropriate

to the species, their developmental stage and behavioural needs.

12.4. Enrichment and handling

Cephalopods shall be provided with the appropriate and

sufficient amount of physical, cognitive and sensory stimuli to enable

a wide range of species specific behaviours. Housing conditions shall

take into account species specific social needs (i.e. group- or solitary-

living habits of the species). Shelters shall be provided, whenever

appropriate for the species.

Where possible, cephalopods shall be handled without taking

them out from the water. Handling of cephalopods in and outside of

the water shall be kept to a minimum and equipment in direct contact

with animals shall be moistened.

Table 12.1

Cephalopods

Group

Body

Minimum

Minimum

Minimum

length

water

water

water depth

surface area

surface area

(cm)

(cm)

for each

(cm 2 )

additional

animal in

group-

holding

(cm 2 )

Cuttlefish

up to 2

>2 to 6

>6 to 12

>12

Sepiolidae

Sepiolids

up to 1

>1 to 3

>3

Loliginidae

Squid

up to 15

>15 to 25

>25

Octopodidae

Octopus

up to 10

>10 to 20

>20

50 " .

3. Annex IV to the principal regulations shall be amended as

(a) item 2 thereof shall be substituted by the following

new item

"2. The killing of animals shall be completed by

one of the following instances

(a) a confirmation of permanent cessation of

the circulation;

(b)

destruction of the brain;

(c)

dislocation of the neck;

(d)

exsanguination; or

(e)

confirmation of the onset of rigor mortis.

Methods of confirmation of death shall be appropriate

for the species to be killed.";

(b)

item 3 thereof shall be amended as follows

(i) the table thereof shall be substituted by the

following new table

"

";

(c) immediately after item 16 thereof there shall be added

the following new item

"17. To be used only for zebrafish (Danio rerio) ≥ 16

days post fertilisation (dpf) and with a maximum body length

of 5cm. Temperature of hypothermic shock shall be ≤ 4°C

and the temperature difference from holding temperature

shall be ≥ 20°C. The fish shall not have direct contact with

the ice. Minimum exposure time shall be 5 minutes.".

€0.81